Identifying human b cells expressing anti&amp;#8208;allergen antibodies

ABSTRACT

In one aspect, methods of generating human monoclonal antibodies that specifically binds to an allergen are provided. In some embodiments, the monoclonal antibodies are generated from sequences identified from isolated single B cells from a human subject who is allergic to the allergen.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of application Ser. No. 17/056,384filed Nov. 17, 2020 (pending), which is the § 371 U.S. National Stage ofPCT Application No. PCT/US2019/032951, filed May 17, 2019, whichpublished under serial no. WO 2019/222679 on Nov. 21, 2019. The PCTapplication claims the priority benefit of U.S. Provisional PatentApplication No. 62/673,713, filed May 18, 2018. The aforesaid priorityapplications are hereby incorporated herein by reference in theirentirety for all purposes.

REFERENCE TO A SEQUENCE LISTING

The Sequence Listing written in file 103182-1134892-000710WO_SL.txtcreated on Nov. 15, 2019, 480 KB, machine format IBM-PC, MS-Windowsoperating system, is hereby incorporated by reference in its entiretyfor all purposes.

BACKGROUND OF THE INVENTION

Allergies are a growing health concern worldwide and are characterizedby a misdirected adaptive immune response towards otherwise harmlessproteins. For food allergies in particular, individuals must be diligentin avoiding allergen exposure or otherwise risk potentially fatalallergic reactions. No cure for allergies exist, and althoughdesensitization regimens such as immunotherapy have shown some clinicalbenefit, there is a need for a fast, effective intervention that canimprove the quality of life for allergic individuals.

BRIEF SUMMARY OF THE INVENTION

In one aspect, the present disclosure provides methods of generating ahuman monoclonal antibody that specifically binds to an allergen. Insome embodiments, the method comprises:

-   -   (a) isolating single B cells from a sample from a human subject,        wherein the subject is allergic to the allergen;    -   (b) generating cDNAs from the single B cells of step (a),        wherein the cDNA sequences comprise a first sequence that        encodes all or part of an immunoglobulin heavy chain and a        second sequence that encodes all or part of an immunoglobulin        light chain;    -   (c) determining the sequences of the cDNAs from step (b);    -   (d) analyzing the sequences determined in step (c) to identify        single B cells comprising a first sequence that comprises an IgE        constant region or an IgG4 constant region;    -   (e) identifying, from the single B cells of step (d), (i) a        heavy chain variable region sequence in the immunoglobulin heavy        chain that comprises an IgE constant region or an IgG4 constant        region, and (ii) a light chain variable region sequence in an        immunoglobulin light chain that is co-expressed with the        immunoglobulin heavy chain in the same single B cell;    -   (f) expressing antibodies comprising the heavy chain variable        region sequence and the light chain variable region sequence        from step (e); and    -   (g) identifying one or more antibodies from step (f) that        specifically bind to the allergen.

In some embodiments, step (a) comprises sorting cells in the sample byfluorescent activated cell sorting (FACS). In some embodiments, step (a)comprises selecting single B cells for expression of cell surface IgEand/or cell surface IgG4. In some embodiments, step (a) comprisesisolating antibody-secreting B cells and/or memory B cells.

In some embodiments, the method comprises isolating single B cells thatare selected for expression of cell surface IgE and identifying asequence encoding an immunoglobulin heavy chain that comprises an IgEconstant region. In some embodiments, step (a) comprises contactingcells from the sample with an anti-human CD19 antibody and an anti-humanIgE antibody and selecting for CD19⁺ IgE-expressing B cells. In someembodiments, the method comprises isolating single B cells that expressa B cell marker and that are negative for non-IgE isotypes. In someembodiments, step (a) comprises contacting cells from the sample with ananti-human CD19 antibody, an anti-human IgM antibody, an anti-human IgGantibody, an anti-human IgA antibody, and an anti-human IgD antibody andselecting for CD19⁺IgM⁻IgG⁻IgA⁻IgD⁻ B cells.

In some embodiments, the method comprises isolating single B cells thatare selected for expression of cell surface IgG4 and identifying asequence encoding an immunoglobulin heavy chain that comprises an IgG4constant region. In some embodiments, step (a) comprises contactingcells from the sample with an anti-human CD19 antibody and an anti-humanIgG4 antibody and selecting for CD19⁺IgG4-expressing B cells. In someembodiments, the method comprises isolating single B cells that expressa B cell marker and that are negative for non-IgG4 isotypes. In someembodiments, step (a) comprises contacting cells from the sample with ananti-human CD19 antibody, an anti-human IgM antibody, an anti-human IgEantibody, an anti-human IgA antibody, an anti-human IgD antibody, ananti-human IgG1 antibody, an anti-human IgG2 antibody, and an anti-humanIgG3 antibody and selecting for CD19⁺IgM⁻IgE⁻IgA⁻IgD⁻IgG1⁻IgG2⁻IgG3⁻ Bcells.

In some embodiments, step (b) comprises reverse transcribing cDNAs fromRNA from the single B cells and amplifying the cDNAs. In someembodiments, the RNA is mRNA. In some embodiments, the method comprisesamplifying immunoglobulin heavy chain and light chain sequences from thesingle B cells. In some embodiments, the method comprises amplifying thetranscriptomes of the single B cells.

In some embodiments, step (f) comprises expressing the heavy chainvariable region sequence and the light chain variable region sequencefrom step (e) in a host cell and purifying the antibodies. In someembodiments, step (f) comprises expressing antibodies comprising theheavy chain variable region sequence and the light chain variable regionsequence from step (e) and an IgG4 constant region or an IgG1 constantregion.

In some embodiments, the method further comprises substituting theconstant region of an antibody identified in step (g) with a wild-typeIgG4 constant region or a modified IgG4 constant region.

In some embodiments, the sample comprises peripheral blood. In someembodiments, the sample comprises tissue (e.g., tonsil tissue).

In some embodiments, the allergen is a food allergen, a plant allergen,a fungal allergen, an animal allergen, a drug allergen, a cosmeticallergen, or a latex allergen. In some embodiments, the allergen is afood allergen selected from the group consisting of a milk allergen, anegg allergen, a nut allergen, a fish allergen, a shellfish allergen, asoy allergen, a legume allergen, a seed allergen, and a wheat allergen.In some embodiments, the food allergen is a peanut allergen. In someembodiments, the food allergen is a tree nut allergen. In someembodiments, the food allergen is a milk allergen. In some embodiments,the allergen is a fungal allergen. In some embodiments, the fungalallergen is an Aspergillus allergen.

In another aspect, monoclonal antibodies produced according to a methoddisclosed herein are provided.

In yet another aspect, pharmaceutical compositions comprising amonoclonal antibody produced according to a method disclosed herein areprovided. In some embodiments, the pharmaceutical composition comprisesa plurality of monoclonal antibodies, wherein each monoclonal antibodyis produced according to a method disclosed herein and wherein themonoclonal antibodies recognize different epitopes or specifically bindto different antigens (e.g., different allergens within the same type orclass of allergen or in different types or classes of allergens).

In still another aspect, monoclonal antibodies, or antigen-bindingportions thereof, that specifically bind to an allergen are provided. Insome embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region sequence that is derived from        an immunoglobulin heavy chain from an IgE- or IgG4-producing        single B cell from a human subject who is allergic to the        allergen;    -   (b) a heavy chain IgG constant region sequence;    -   (c) a light chain variable region sequence that is derived from        an immunoglobulin light chain from the IgE-producing or        IgG4-producing single B cell from a human subject; and    -   (d) a light chain constant region sequence that is of the same        class as the immunoglobulin light chain of (c).

In some embodiments, the antibody or the antigen-binding portion thereofspecifically binds to an allergen with a binding affinity (K_(D)) ofless than 1 nM. In some embodiments the antibody or the antigen-bindingportion thereof specifically binds to an allergen with a bindingaffinity (KD) of less than 250 nM, less than 100 nM, less than 50 nM,less than 10 nM, or less than 5 nM. In some embodiments, the antibodybinds to the allergen with a binding affinity (KD) from 1 nM to 250 nM.In some embodiments, the allergen is a food allergen, a plant allergen,a fungal allergen, an animal allergen, a dust mite allergen, a drugallergen, a cosmetic allergen, or a latex allergen. In some embodiments,the allergen is a food allergen selected from the group consisting of amilk allergen, an egg allergen, a nut allergen, a fish allergen, ashellfish allergen, a soy allergen, a legume allergen, a seed allergen,and a wheat allergen. In some embodiments, the food allergen is a peanutallergen or a tree nut allergen. In some embodiments, the food allergenis a milk allergen. In some embodiments, the allergen is a fungalallergen. In some embodiments, the fungal allergen is an Aspergillusantigen. In some embodiments, the antibody is cross-reactive with twodifferent antigens. In some embodiments, the antibody is cross-reactivewith a first antigen of a first allergen type and a second antigen of asecond allergen type that is different from the first allergen type. Insome embodiments, the antibody is cross-reactive with a peanut allergenand a tree nut allergen. In some embodiments, the antibody iscross-reactive with two or more tree nut antigens.

In some embodiments, the antibody comprises a heavy chain variableregion sequence and a light chain variable region sequence that arederived from an IgE-producing human B cell or from an IgG4-producinghuman B cell.

In yet another aspect, monoclonal antibodies, or antigen-bindingportions thereof, that specifically binds to a peanut allergen areprovided. In some embodiments, the antibody binds to the peanut allergenwith a binding affinity (K_(D)) of less than 1 nM. In some embodimentsthe antibody or the antigen-binding portion thereof specifically bindsto an peanut allergen with a binding affinity (KD) of less than 250 nM,less than 100 nM, less than 50 nM, less than 10 nM, or less than 5 nM.In some embodiments, the antibody binds to the peanut allergen with abinding affinity (KD) from 1 nM to 250 nM.

In some embodiments, the antibody or the antigen-binding portion thereofspecifically binds to the peanut allergen Ara h 2, Ara h 3, or Ara h 1.In some embodiments, the antibody or the antigen-binding portion thereofspecifically binds to Ara h 2 with a K_(D) of less than 100 pM. In someembodiments, the antibody or the antigen-binding portion thereof iscross-reactive with at least two peanut allergens. In some embodiments,the antibody or the antigen-binding portion thereof is cross-reactivewith Ara h 2 and Ara h 3. In some embodiments, the antibody or theantigen-binding portion thereof specifically binds to Ara h 2 with aK_(D) of less than 1 nM and specifically binds to Ara h 3 with a K_(D)of less than 1 μM. In some embodiments, the antibody or theantigen-binding portion thereof binds to an epitope that comprises theamino acid motif DPYSPS (SEQ ID NO:704). Furthermore, additionalpeanut-specific antibodies were discovered during these experiments.Antibodies PA12P3E09 and PA12P3E11 bound peanut extract with little tono binding to natural peanut allergen Ara h 2, while antibodiesPA12P1D02, PA12P1G11, PA13P1H03, PA12P3C01, and PA12P3E04 bound stronglyto both peanut extract and natural peanut allergen Ara h 2. In someembodiments the peanut specific antibody binds to peanut extract, butdoes not bind natural peanut allergen Ara h 2.

In some embodiments, the antibody comprises:

-   -   a heavy chain CDR1 comprising the amino acid sequence of any one        of SEQ ID NOs:2, 10, 17, 25, 33, 41, 47, 58, 113, 129, 199, 341,        348, 409, 459, or 593;    -   a heavy chain CDR2 comprising the amino acid sequence of any one        of SEQ ID NOs:3, 11, 18, 26, 34, 48, 59, 130, 200, 342, 349,        410, 460, 539, or 594;    -   a heavy chain CDR3 comprising the amino acid sequence of any one        of SEQ ID NOs:4, 12, 19, 27, 35, 42, 49, 55, 60, 131, 201, 350,        411, 461, 540, or 595;    -   a light chain CDR1 comprising the amino acid sequence of any one        of SEQ ID NOs:6, 14, 21, 29, 37, 44, 51, 62, 133, 203, 343, 352,        413, 463, 542, or 597;    -   a light chain CDR2 comprising the amino acid sequence of any one        of SEQ ID NOs:7, 15, 22, 30, 38, 52, 78, 86, 126, 149, 196, 345,        353, or 598; and    -   a light chain CDR3 comprising the amino acid sequence of any one        of SEQ ID NOs:8, 23, 31, 39, 45, 53, 63, 134, 204, 346, 354,        414, 464, 543, or 599.

In some embodiments, the antibody comprises heavy chain and light chainCDR sequences contained within the heavy chain variable region and lightchain variable region sequence pairs selected from the group consistingof SEQ ID NOs: 1 and 5; 9 and 13; 16 and 20; 24 and 28; 32 and 36; 40and 43; 46 and 50; 54 and 56; 57 and 61; 57 and 5; 1 and 61; 64 and 5;65 and 5; 66 and 5; 67 and 5; 128 and 132; 340 and 344; 347 and 351; 406and 407; 408 and 412; 458 and 462; 538 and 541; and 592 and 596. In someembodiments, the antibody comprises a heavy chain CDR1-3 and a lightchain CDR1-3 comprising the amino acid sequences of:

-   -   (a) SEQ ID NOs: 2, 3, 4, 6, 7, and 8, respectively; or    -   (b) SEQ ID NOs: 10, 11, 12, 14, 15, and 8, respectively; or    -   (c) SEQ ID NOs: 17, 18, 19, 21, 22, and 23, respectively; or    -   (d) SEQ ID NOs: 25, 26, 27, 29, 30, and 31, respectively; or    -   (e) SEQ ID NOs: 33, 34, 35, 37, 38, and 39, respectively; or    -   (f) SEQ ID NOs: 41, 34, 35, 37, 38, and 39, respectively; or    -   (g) SEQ ID NOs: 47, 48, 49, 51, 52, and 53, respectively; or    -   (h) SEQ ID NOs: 47, 48, 55, 51, 52, and 53, respectively; or    -   (i) SEQ ID NOs: 58, 59, 60, 62, 30, and 63, respectively; or    -   (j) SEQ ID NOs: 58, 59, 60, 6, 7, and 8, respectively; or    -   (k) SEQ ID NOs: 2, 3, 4, 62, 30, and 63, respectively; or    -   (l) SEQ ID NOs: 58, 3, 4, 6, 7, and 8, respectively; or    -   (m) SEQ ID NOs: 2, 59, 4, 6, 7, and 8, respectively; or    -   (n) SEQ ID NOs: 2, 3, 60, 6, 7, and 8, respectively; or    -   (o) SEQ ID NOs:129, 130, 131, 133, 126, and 134, respectively;        or    -   (p) SEQ ID NOs:341, 342, 343, 345, 78, and 346, respectively; or    -   (q) SEQ ID NOs:348, 349, 350, 352, 353, and 354, respectively;        or    -   (r) SEQ ID NOs:199, 200, 201, 203, 149, and 204, respectively;        or    -   (s) SEQ ID NOs:409, 410, 411, 413, 86, and 414, respectively; or    -   (t) SEQ ID NOs:459, 460, 461, 463, 196, and 464, respectively;        or    -   (u) SEQ ID NOs:113, 539, 540, 542, 196, and 543, respectively;        or    -   (v) SEQ ID NOs:593, 594, 595, 597, 598, and 599, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity to any one of SEQ ID NOs:1, 9, 16, 24, 32, 40, 46, 54, 57, 64,65, 66, 67, 128, 340, 347, 406, 408, 458, 538, or 592. In someembodiments, the antibody comprises a light chain variable regioncomprising an amino acid sequence that has at least 90% sequenceidentity to any one of SEQ ID NOs:5, 13, 20, 28, 36, 43, 50, 56, 61,132, 344, 351, 407, 412, 462, 541, or 596.

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID NO:1        and a light chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID NO:5;        or    -   (b) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID NO:9        and a light chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:13; or    -   (c) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID NO:16        and a light chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:20; or    -   (d) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID NO:24        and a light chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:28; or    -   (e) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID NO:32        and a light chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:36; or    -   (f) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID NO:40        and a light chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:43; or    -   (g) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID NO:46        and a light chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:50; or    -   (h) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID NO:54        and a light chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:56; or    -   (i) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID NO:57        and a light chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:61; or    -   (j) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID NO:57        and a light chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID NO:5;        or    -   (k) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID NO:1        and a light chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:61; or    -   (l) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID NO:64        and a light chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID NO:5;        or    -   (m) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID NO:65        and a light chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID NO:5;        or    -   (n) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID NO:66        and a light chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID NO:5;        or    -   (o) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID NO:67        and a light chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID NO:5;        or    -   (p) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:128 and a light chain variable region comprising an amino        acid sequence that has at least 90% sequence identity to SEQ ID        NO:132; or    -   (q) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:340 and a light chain variable region comprising an amino        acid sequence that has at least 90% sequence identity to SEQ ID        NO:344; or    -   (r) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:347 and a light chain variable region comprising an amino        acid sequence that has at least 90% sequence identity to SEQ ID        NO:351; or    -   (s) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:406 and a light chain variable region comprising an amino        acid sequence that has at least 90% sequence identity to SEQ ID        NO:407; or    -   (t) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:408 and a light chain variable region comprising an amino        acid sequence that has at least 90% sequence identity to SEQ ID        NO:412; or    -   (u) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:458 and a light chain variable region comprising an amino        acid sequence that has at least 90% sequence identity to SEQ ID        NO:462; or    -   (v) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:538 and a light chain variable region comprising an amino        acid sequence that has at least 90% sequence identity to SEQ ID        NO:541; or    -   (w) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:592 and a light chain variable region comprising an amino        acid sequence that has at least 90% sequence identity to SEQ ID        NO:596.

In still another aspect, monoclonal antibodies, or antigen-bindingportions thereof, that specifically binds to a tree nut allergen areprovided. In some embodiments, the antibody binds to the tree nutallergen with a binding affinity (K_(D)) of less than 250 nM, less than100 nM, less than 50 nM, less than 10 nM, or less than 5 nM. In someembodiments, the antibody binds to the tree nut allergen with a bindingaffinity (K_(D)) of less than 1 nM. In some embodiments, the antibodybinds to the tree nut allergen with a binding affinity (K_(D)) from 1 nMto 250 nM. In some embodiments, the tree nut allergen is cashew,pistachio, pecan, walnut, hazelnut, and/or macadamia nut.

In some embodiments, the antibody comprises:

-   -   a heavy chain CDR1 comprising the amino acid sequence of any one        of SEQ ID NOs:113, 167, 175, 227, 311, 318, 438, 466, 621, 665,        or 692;    -   a heavy chain CDR2 comprising the amino acid sequence of any one        of SEQ ID NOs: 168, 176, 200, 312, 319, 439, 539, 666, or 693;    -   a heavy chain CDR3 comprising the amino acid sequence of any one        of SEQ ID NOs: 169, 177, 228, 313, 320, 440, 467, 540, 667, or        694;    -   a light chain CDR1 comprising the amino acid sequence of any one        of SEQ ID NOs: 171, 179, 230, 315, 322, 442, 469, 542, 623, 669,        or 696;    -   a light chain CDR2 comprising the amino acid sequence of any one        of SEQ ID NOs: 30, 94, 149, 172, 180, 196, 323, or 670; and    -   a light chain CDR3 comprising the amino acid sequence of any one        of SEQ ID NOs: 173, 181, 231, 316, 324, 443, 470, 543, 624, 671,        or 697.

In some embodiments, the antibody comprises a heavy chain CDR1-3 and alight chain CDR1-3 comprising the amino acid sequences of:

-   -   (a) SEQ ID NOs: 692, 693, 694, 696, 94, and 697, respectively;        or    -   (b) SEQ ID NOs:318, 319, 320, 322, 323, and 324, respectively;        or    -   (c) SEQ ID NOs:227, 200, 228, 230, 149, and 231, respectively;        or    -   (d) SEQ ID NOs:113, 539, 540, 542, 196, and 543, respectively;        or    -   (e) SEQ ID NOs:311, 312, 313, 315, 94, and 316, respectively; or    -   (f) SEQ ID NOs:665, 666, 667, 669, 670, and 671, respectively;        or    -   (g) SEQ ID NOs:466, 200, 467, 469, 149, and 470, respectively;        or    -   (h) SEQ ID NOs:167, 168, 169, 171, 172, and 173, respectively;        or    -   (i) SEQ ID NOs:621, 176, 177, 623, 180, and 624, respectively;        or    -   (j) SEQ ID NOs:175, 176, 177, 179, 180, and 181, respectively;        or    -   (k) SEQ ID NOs:438, 439, 440, 442, 30, and 443, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity to any one of SEQ ID NOs:166, 174, 226, 310, 317, 437, 465,538, 620, 664, or 691. In some embodiments, the antibody comprises alight chain variable region comprising an amino acid sequence that hasat least 90% sequence identity to any one of SEQ ID NOs:170, 178, 229,314, 321, 441, 468, 541, 622, 668, or 695.

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:691 and a light chain variable region comprising an amino        acid sequence that has at least 90% sequence identity to SEQ ID        NO:695; or    -   (b) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:317 and a light chain variable region comprising an amino        acid sequence that has at least 90% sequence identity to SEQ ID        NO:321; or    -   (c) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:226 and a light chain variable region comprising an amino        acid sequence that has at least 90% sequence identity to SEQ ID        NO:229; or    -   (d) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:538 and a light chain variable region comprising an amino        acid sequence that has at least 90% sequence identity to SEQ ID        NO:541; or    -   (e) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:310 and a light chain variable region comprising an amino        acid sequence that has at least 90% sequence identity to SEQ ID        NO:314; or    -   (f) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:664 and a light chain variable region comprising an amino        acid sequence that has at least 90% sequence identity to SEQ ID        NO:668; or    -   (g) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:465 and a light chain variable region comprising an amino        acid sequence that has at least 90% sequence identity to SEQ ID        NO:468; or    -   (h) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:166 and a light chain variable region comprising an amino        acid sequence that has at least 90% sequence identity to SEQ ID        NO:170;

or

-   -   (i) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:620 and a light chain variable region comprising an amino        acid sequence that has at least 90% sequence identity to SEQ ID        NO:622; or    -   (j) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:174 and a light chain variable region comprising an amino        acid sequence that has at least 90% sequence identity to SEQ ID        NO:178; or    -   (k) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:437 and a light chain variable region comprising an amino        acid sequence that has at least 90% sequence identity to SEQ ID        NO:441.

In yet another aspect, monoclonal antibodies, or antigen-bindingportions thereof, that specifically binds to a milk allergen areprovided. In some embodiments, the antibody binds to the milk allergenwith a binding affinity (KD) of less than 250 nM, less than 100 nM, lessthan 50 nM, less than 10 nM, or less than 5 nM. In some embodiments, theantibody binds to the milk allergen with a binding affinity (K_(D)) ofless than 1 nM. In some embodiments, the antibody binds to the milkallergen with a binding affinity (KD) from 1 nM to 250 nM.

In some embodiments, the antibody comprises:

-   -   a heavy chain CDR1 comprising the amino acid sequence of any one        of SEQ ID NOs: 121, 750, 757, 765, 772, 779, 785, 793, 800, 807,        814, 821, 833, 838, 846, 853, 860, 868, 874, 881, 889, 895, 903,        911, 918, or 926;    -   a heavy chain CDR2 comprising the amino acid sequence of any one        of SEQ ID NOs: 378, 532, 751, 758, 766, 773, 786, 794, 801, 808,        815, 822, 826, 839, 847, 854, 861, 875, 882, 890, 896, 904, 912,        919, or 927;    -   a heavy chain CDR3 comprising the amino acid sequence of any one        of SEQ ID NOs: 752, 759, 767, 774, 780, 787, 795, 802, 809, 816,        827, 840, 848, 855, 862, 869, 876, 883, 891, 897, 905, 913, 920,        or 928;    -   a light chain CDR1 comprising the amino acid sequence of any one        of SEQ ID NOs: 523, 754, 761, 769, 776, 782, 789, 797, 804, 811,        818, 829, 835, 842, 850, 857, 864, 871, 878, 885, 899, 907, 915,        922, or 930;    -   a light chain CDR2 comprising the amino acid sequence of any one        of SEQ ID NOs: 22, 30, 94, 110, 149, 186, 196, 389, 404, 509,        662, 682, 762, 790, 830, 843, 865, 886, 900, 908, 923, or 931;        and    -   a light chain CDR3 comprising the amino acid sequence of any one        of SEQ ID NOs: 755, 763, 770, 777, 783, 791, 798, 805, 812, 819,        824, 831, 836, 844, 851, 858, 866, 872, 879, 887, 893, 901, 909,        916, 924, or 932.

In some embodiments, the antibody comprises a heavy chain CDR1-3 and alight chain CDR1-3 comprising the amino acid sequences of:

-   -   (a) SEQ ID NOs: 800, 801, 802, 804, 110, and 805, respectively;        or    -   (b) SEQ ID NOs:121, 826, 827, 829, 830, and 831, respectively;        or    -   (c) SEQ ID NOs:833, 826, 827, 835, 149, and 836, respectively;        or    -   (d) SEQ ID NOs:853, 854, 855, 857, 662, and 858, respectively;        or    -   (e) SEQ ID NOs:860, 861, 862, 864, 865, and 866, respectively;        or    -   (f) SEQ ID NOs:868, 378, 869, 871, 682, and 782, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity to any one of SEQ ID NOs:749, 756, 764, 771, 778, 784, 792,799, 806, 813, 820, 825, 832, 837, 845, 852, 859, 867, 873, 880, 888,894, 902, 910, 917, or 925. In some embodiments, the antibody comprisesa light chain variable region comprising an amino acid sequence that hasat least 90% sequence identity to any one of SEQ ID NOs:753, 760, 768,775, 781, 788, 796, 803, 810, 817, 823, 828, 834, 841, 849, 856, 863,870, 877, 884, 892, 898, 906, 914, 921, or 929.

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:799 and a light chain variable region comprising an amino        acid sequence that has at least 90% sequence identity to SEQ ID        NO:803; or    -   (b) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:825 and a light chain variable region comprising an amino        acid sequence that has at least 90% sequence identity to SEQ ID        NO:828; or    -   (c) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:832 and a light chain variable region comprising an amino        acid sequence that has at least 90% sequence identity to SEQ ID        NO:834; or    -   (d) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:852 and a light chain variable region comprising an amino        acid sequence that has at least 90% sequence identity to SEQ ID        NO:856; or    -   (e) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:859 and a light chain variable region comprising an amino        acid sequence that has at least 90% sequence identity to SEQ ID        NO:863; or    -   (f) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:867 and a light chain variable region comprising an amino        acid sequence that has at least 90% sequence identity to SEQ ID        NO:870.

In some embodiments, the antibody is an antibody that competes with amonoclonal antibody as disclosed herein for binding to an allergen(e.g., for binding to a food allergen such as a peanut allergen, a treenut allergen, or a milk allergen).

In yet another aspect, monoclonal antibodies, or antigen-bindingportions thereof, that specifically binds to a fungal allergen areprovided. In some embodiments, the fungal allergen is an Aspergillusallergen. In some embodiments, the antibody binds to the fungal allergen(e.g., Aspergillus allergen) with a binding affinity (KD) of less than250 nM, less than 100 nM, less than 50 nM, less than 10 nM, or less than5 nM. In some embodiments, the antibody binds to the fungal allergen(e.g., Aspergillus allergen) with a binding affinity (K_(D)) of lessthan 1 nM. In some embodiments, the antibody binds to the fungalallergen with a binding affinity (KD) from 1 nM to 250 nM.

In some embodiments, the antibody or the antigen-binding portion thereofspecifically binds to the allergen Aspergillus fumigatus, Aspergillusniger, and/or Aspergillus nidulans. In some embodiments, the antibodyspecifically binds to the allergen Aspergillus fumigatus 1 (Asp f 1).

In some embodiments, the antibody comprises:

-   -   a heavy chain CDR1 comprising the amino acid sequence of any one        of SEQ ID NOs:710, 718, 726, 734, or 742;    -   a heavy chain CDR2 comprising the amino acid sequence of any one        of SEQ ID NOs:711, 719, 727, 735, or 743;    -   a heavy chain CDR3 comprising the amino acid sequence of any one        of SEQ ID NOs:712, 720, 728, 736, or 744;    -   a light chain CDR1 comprising the amino acid sequence of any one        of SEQ ID NOs:714, 722, 730, 738, or 746;    -   a light chain CDR2 comprising the amino acid sequence of any one        of SEQ ID NOs:715, 723, 731, 739, or 747; and    -   a light chain CDR3 comprising the amino acid sequence of any one        of SEQ ID NOs:716, 724, 732, 740, or 748.

In some embodiments, the antibody comprises heavy chain and light chainCDR sequences contained within the heavy chain variable region and lightchain variable region sequence pairs selected from the group consistingof SEQ ID NOs:709 and 713; 717 and 721; 725 and 729; 733 and 737; and741 and 745. In some embodiments, the antibody comprises a heavy chainCDR1-3 and a light chain CDR1-3 comprising the amino acid sequences of:

-   -   (a) SEQ ID NOs: 710, 711, 712, 714, 715, and 716, respectively;        or    -   (b) SEQ ID NOs:718, 719, 720, 722, 723, and 724, respectively;        or    -   (c) SEQ ID NOs:726, 7272, 728, 730, 731, and 732, respectively;        or    -   (d) SEQ ID NOs:734, 735, 736, 738, 739, and 740, respectively;        or    -   (e) SEQ ID NOs:742, 743, 744, 746, 747, and 748, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity to any one of SEQ ID NOs:709, 717, 725, 733, or 741. In someembodiments, the antibody comprises a light chain variable regioncomprising an amino acid sequence that has at least 90% sequenceidentity to any one of SEQ ID NOs:713, 721, 729, 737, or 745.

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:709 and a light chain variable region comprising an amino        acid sequence that has at least 90% sequence identity to SEQ ID        NO:713; or    -   (b) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:717 and a light chain variable region comprising an amino        acid sequence that has at least 90% sequence identity to SEQ ID        NO:721; or    -   (c) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:725 and a light chain variable region comprising an amino        acid sequence that has at least 90% sequence identity to SEQ ID        NO:729; or    -   (d) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:733 and a light chain variable region comprising an amino        acid sequence that has at least 90% sequence identity to SEQ ID        NO:737; or    -   (e) a heavy chain variable region comprising an amino acid        sequence that has at least 90% sequence identity to SEQ ID        NO:741 and a light chain variable region comprising an amino        acid sequence that has at least 90% sequence identity to SEQ ID        NO:745.

In some embodiments, the antibody is an antibody that competes with amonoclonal antibody as disclosed herein for binding to a fungal allergen(e.g., for binding to an Aspergillus allergen).

In another aspect, pharmaceutical compositions comprising a monoclonalantibody or antigen-binding portion as disclosed herein are provided. Insome embodiments, the pharmaceutical composition comprises a pluralityof monoclonal antibodies as disclosed herein, wherein the monoclonalantibodies recognize different epitopes or specifically bind todifferent antigens (e.g., different allergens within the same type orclass of allergen or in different types or classes of allergens).

In another aspect, antibody-drug conjugates comprising a monoclonalantibody or antigen-binding portion thereof as disclosed herein areprovided. In some embodiments, the antibody-drug conjugate comprises amonoclonal antibody or antigen-binding portion thereof that specificallybinds to a fungal allergen as disclosed herein and further comprises ananti-fungal agent. In some embodiments, the anti-fungal agent isAmphotericin B.

In still another aspect, isolated polynucleotides comprising anucleotide sequence encoding a monoclonal antibody as disclosed herein.Also provided herein are vectors and host cells comprising apolynucleotide as disclosed herein.

In another aspect, therapeutic methods are provided. In someembodiments, the therapeutic method is a method of reducing one or moreallergy symptoms in a subject. In some embodiments, the therapeuticmethod is a method of reducing one or more allergy symptoms in a subjecthaving a peanut allergy. In some embodiments, the therapeutic method isa method of reducing one or more allergy symptoms in a subject having atree nut allergy. In some embodiments, the therapeutic method is amethod of reducing one or more allergy symptoms in a subject having afungal allergy. In some embodiments, the therapeutic method is a methodof reducing one or more allergy symptoms in a subject having a milkallergy. In some embodiments, the method comprises administering to thesubject a therapeutically effective amount of a monoclonal antibody orpharmaceutical composition as disclosed herein.

In another aspect, kits are provided. In some embodiments, the kitcomprises a monoclonal antibody or pharmaceutical composition asdisclosed herein. In some embodiments, the kit is for use in practicinga therapeutic method as disclosed herein.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1. Flowchart of exemplary workflow for method of generatingallergen-specific monoclonal antibodies.

FIGS. 2A-2G. Characterization of single B cells isolated from freshhuman peripheral blood. (A) Principal component analysis (left)separates naive/memory (black dots) and plasmablast (PB, gray dots) Bcell subsets identified by expression of established transcriptionfactors and marker genes (right; non-striped for naive/memory B cellsand striped for plasmablast (PB) B cells). (B) Isotype of B cells, blackdots for IGHE and gray dots for other isotypes. (C) Number of cellsbelonging to each subtype in (A) by isotype (non-striped fornaive/memory B cells and striped for plasmablast (PB) B cells). (D-G)Analysis of clonal families (CFs). (D) Distribution of the number ofcells per CF. (E) Fraction of cells of each isotype that belong to amulti-member CF. (F) Isotype, B cell subtype, mutational frequency, andpatient of origin of each cell within multi-member CFs. CFs referred toin the text are labeled. (G) Heavy (right) and light (left) chain CDR3sequences and similarity heatmap for CFs in (F).

FIGS. 3A-3G. Characterization of 89 IgE antibodies from single B cells.(A) Phylogenetic depiction of antibody heavy chains arranged by IGHVgene (background pattern), patient of origin (node pattern), andmutation frequency (node size). (B) Heatmap indicating number of cellswith a given heavy and light chain CDR3 length. (C) Heavy and lightchain mutation frequency of each cell. (D) Silent (S) and replacement(R) mutations by region within heavy and light chains. (E) Differentialgene expression between IgE PBs and PBs of other isotypes. Positive logfold change indicates genes enriched in IgE PBs. (F) Heavy chainconstant region coverage histograms for naïve/memory B cells (top) andPBs (bottom) by isotype, with loci oriented in the 5′ to 3′ direction.Mean normalized read depth and 95% confidence interval are indicated bysolid lines and shaded area, respectively, for the number of cells (n)inscribed. Membrane exons are the two most 3′ exons of each isotype. (G)Summary of (F), but depicting the fraction of cells of each isotype withany membrane exon coverage.

FIGS. 4A-4C. High affinity, cross-reactive IgE antibody convergence intwo unrelated individuals (PA12 & PA13). Antibody patterns are conservedamong panels. (A) Frequency of silent (S) and replacement (R) mutationsby region. (B) Mutation frequency percentiles compared to allclass-switched antibodies. (C) Dissociation constants (KDs) for majorallergenic peanut proteins Ara h 2 and Ara h 3 for six convergentantibodies as well as eight variants of PA13P1H08. Shortened antibodyvariant names are designated as “heavy-light,” using the followingabbreviations: N=native, R=reverted, FWRs=framework regions. An “r”prefix indicates only that region has been reverted.

FIGS. 5A-5E. Germline transcription reveals class switch priming insingle B cells. (A) Identity of CE germline transcript splice donorsalong with the number of cells, by isotype, expressing each. (B)Fraction of cells expressing EGLTs by isotype. (C) Example fromindividual PA11 where identification of phased variants within IgEconstant region exons enables subsequent verification of biallelic GLTexpression in other B cells from the same patient. (D) Global germlinetranscription state heatmap indicating the fraction of cells of a givenisotype expressing a given GLT. Above: GLT isotype expression frequencyrelative to all GLT isotypes; excludes self-isotype GLT expression. (E)Histogram of the number of non-self GLT isotypes expressed in each cell.

FIG. 6. Study overview. Plasma was extracted from fresh blood to measurecirculating IgE levels, while the cellular fraction was enriched for Bcells prior to FACS. Single cells were sorted into individual wells of a96 well plate and processed with scRNA-seq, generating sequencing readsthat were aligned to the genome to calculate gene expression and assesssplicing as well as assembled in order to reconstruct heavy and lightchain sequences. Specificity and affinity data were generated forrecombinantly expressed antibodies.

FIGS. 7A-7C. Plasma IgE levels. (A) Allergen-specific and allergencomponent (hazelnut, peanut) concentrations. (B) Total IgEconcentration. (C) Positive correlation between total plasma IgE leveland the number confirmed IgE+ B cells. Each point is an individual.

FIGS. 8A-8C. FACS gating and analysis. (A) Gating strategy for sortingsingle B cells. IgE+ B cells have been overlaid as dark gray dots. (B)Isotype identity within the final IgE gate as determined by heavy chaintranscript assembly. ND=not determined. (C) For reference, putativebasophils (CD19−IgE+) display higher IgE surface expression than IgE+ Bcells.

FIGS. 9A-9G. Single cell RNA-seq data overview and quality control. (A)Cells were sequenced in 5 libraries to a depth of ^(˜)1-2 millionreads/cell. (B) Genes per cell histogram. Cells expressing fewer than950 genes were discarded. (C) Rarefaction curve depicting the number ofgenes detected as a function of sequencing depth for eight randomlyselected cells in each B cell subtype. Solid lines and shaded arearepresent mean and 95% confidence interval for the gene count,respectively. (D) Read mapping distribution for retained cells. Mostreads mapped uniquely (Ensembl reference annotation) and multimappedreads largely belonged to RNA18S5 repeats on chr21 and unplacedscaffolds. (E) Read mapping across gene bodies showed minimal 3′ or 5′bias. (F) V gene assembly length histogram by chain. (G) PCA on the top500 most variable genes before (top) and after (bottom) batchcorrection.

FIGS. 10A-10D. Auxiliary data supporting B cell subtype classification.PBs (striped) have greater FACS forward and side scatter (A), more cDNAafter SmartSeq2 preamplification (B), and have greater gene expressionof antibody light and heavy chains (C) as compared to the naive/memory Bcell subset (non-striped). (D) Top differentially expressed genes foreach subset.

FIGS. 11A-11K. B cell comparisons across isotypes. (A-I) Number of cellswith a given V and J gene by isotype and chain. (A) IGHM. (B) IGHD. (C)IGHG3. (D) IGHG1. (E) IGHA1. (F) IGHG2. (G) IGHG4. (H) IGHE. (I) IGHA2.(J) Heavy chain mutation frequency by isotype. (K) Relative utilizationof the lambda and kappa light chain by isotype.

FIGS. 12A-12H. Antibody specificity and affinity characterizations. (A)Semi-quantitative indirect ELISAs of convergent antibodies, controls,PA13P1H08 variants, and antibodies from other clonal families. Human IgGisotype control (abcam #ab206195) served as a negative control, whilepositive controls included anti-Ara h mouse monoclonal antibodiespurchased from Indoor Biotechnologies. (B-G) Kinetic characterization ofantibody binding to Ara h 2 (B-D) and Ara h 3 (E-G) using biolayerinterferometry. Antibodies are named and described herein. (H) IndirectELISA showing binding of recombinant monoclonal antibodies from subjectsPA11, PA12, PA13, PA14, PA15, and PA16 (rows) to allergen extracts,natural peanut allergen Ara h 2, and bovine serum albumin (BSA)(columns). The isotype of each antibody is shown to the left. Highervalues indicate stronger binding. OD=optical density. Depicted valuesrepresent those after subtraction of human IgG isotype control. Only thetested antibodies with any OD value above 0.25 are shown.

FIGS. 13A-13E. Sequences of heavy and light chains used in constructingPA13P1H08 antibody variants. (A) Derivation of the inferred naïve heavychain CDR3 and surrounding amino acids. (B) Native and reverted heavychain sequences, in addition to sequences where region(s) of the heavychain have been reverted to the inferred naïve rearrangement. Labelswith an “r” prefix indicate only that region has been reverted.FWRs=frameworks. (C) Derivation of the inferred naïve light chain CDR3and surrounding amino acids. (D) Native and reverted light chainsequences. (E) Sequence of a light chain taken from another antibody,PA12P4H03, which we confirmed did not to bind any peanut allergens byELISA.

FIG. 14. GLT splicing for all isotypes. Note that only the first threeconstant region exons are shown for each isotype for clarity.

FIG. 15. IGV coverage histograms and splice junctions for the igheconstant region locus showing EGLTs in single murine B cells stimulatedwith IL-4, LPS, and BAFF (Wu et al. 2017). Arrows indicate ighe GLTsplice donors.

FIG. 16. Pairwise CDR3 sequence identity of the heavy chain CDR3sequences from clones PA12P3F10, PA12P3D08, PA12P1C07, PA13P1E10,PA13P3G09, and PA13P1H08 (SEQ ID NOs: 19, 35, 42, 12, 27, 4) and threeheavy chain CDR3 sequences derived from multiple patients in a separatepeanut allergy immune repertoire sequencing study (62). Each sequencefrom this separate study has an identity of at least 70% with one ormore sequences from the present study. All sequences share the IGHV3-30and IGHJ6 gene segments and have CDR3s 17 amino acids in length.

FIG. 17A. Microarray scan confirming the absence of backgroundinteractions of the secondary goat anti-human IgG and the control mousemonoclonal anti-HA antibody with antigen-derived peptides. The controlantibody gave rise to the expected HA control spot pattern framing thepeptide microarray (white dots).

FIGS. 17B and 17C. Microarray epitope mapping of PA13P1H08 to Ara h 2(“Ah2”) and Ara h 3 (“Ah3”) peptides. (B) Microarray scan illustratingantibody binding to antigen peptides (light gray) as well as theexpected HA control spot pattern (white dots). (C) Microarrayfluorescence intensity by antigen peptide. Ara h 2 motifs with highintensity are annotated.

FIG. 18. Plasma IgE levels for milk allergic subject PA01 andAspergillus study subject 10033201 against common food allergens as wellas Aspergillus fumigatus and Aspergillus niger. NP=not performed. TotalIgE was 353 kU/L and 3528 kU/L for PA01 and 10033201, respectively. Theassay was performed by CLIA-licensed Johns Hopkins UniversityDermatology, Allergy, and Clinical Immunology Reference Laboratory usingthe ImmunoCAP system.

FIG. 19. Indirect ELISA showing binding of recombinant monoclonalantibodies (columns) to antigens (rows). Antigens include extracts ofAspergillus fumigatus, Aspergillus niger, and Aspergillus nidulans, aswell as a purified recombinant allergen Aspergillus fumigatus 1 (rAsp f1). Bovine serum albumin (BSA) serves as a negative antigen control. Amonoclonal antibody against Asp f 1 (“anti-Asp f 1”) serves as apositive control against this allergen. Higher values indicate strongerbinding. OD=optical density. hIgG=human IgG isotype control.

FIG. 20. Indirect ELISA showing binding of recombinant monoclonalantibodies from subject PA01 (rows) to allergen extracts, natural peanutallergen Ara h 2, and BSA (columns). The isotype of each antibody isshown to the left. Higher values indicate stronger binding. OD=opticaldensity. Depicted values represent those after subtraction of human IgGisotype control.

TABLE 1

Table 1 includes protein and nucleic acid sequences discussed herein.Polypeptide sequences are provided using the standard one-letter code.One of ordinary skill in the art provided with an amino acid sequencewill understand that the amino acid sequence may be encoded by a definedset of nucleotide sequences such that the reader and inventors haveposession of the nucleotide sequences encoding each amino acid sequence.A nucleic acid sequence encoding a polynucleotide may be a naturallyoccurring human sequence. In some embodiments a nucleic acid sequenceencoding a polynucleotide is not a naturally occurring human sequence. Anucleic acid sequence encoding a polynucleotide may be a sequence thatis codon optimized for expression in human cells or specific cell types,eukaryotic cells, bacteria cells, or otherwise. Codon optimization,which replaces one codon by another codon encoding the same amino acidand having a higher frequency of occurrence in the particular host cell,can be performed to improve the ability of the host to produce thepolypeptide encoded by the nucleic acid (see, e.g., Mauro, BioDrugs32(1):69-81, 2018 and Kato, Intl Mol Sci. 20(4), 2019).

In certain embodiments it is contemplated that variant sequences may beused in methods and compositions disclosed herein. For example, in oneaspect, an antibody with a heavy chain variable region comprising anamino acid sequence that has at least 90% sequence identity to SEQ IDNO:1 and a light chain variable region comprising an amino acid sequencethat has at least 90% sequence identity to SEQ ID NO:5 is described. Adegree of sequence identity or similarity can be determined usingart-known methods. In one approach, the identity of two nucleotide orpolypeptide sequences or subsequences is calculated as the percentage ofpositions that are identical or equivalent after the sequences have beenaligned, introducing gaps, if necessary, to achieve maximum percentsequence identity. Methods of sequence alignment are art-known methods,and include, but are not limited to the Needleman-Wunsch globalalignment algorithm (Needleman and Wunsch (1970) J. Mol. Biol.48(3):443-453 (30)), the Smith and Waterman local homology searchalgorithm (Smith, Temple F. & Waterman, Michael S. (1981) J. Mol. Biol.147 (1): 195-197.), manual alignment and inspection, or computerizedimplementations of these algorithms, such as the “needle” program,distributed as part of the EMBOSS software package (Rice, P. et al.,Trends in Genetics 16(6): 276-277 (31), versions 6.3.1 available fromEMBnet at various sources).

It is contemplated that, in certain embodiments, a method or compositiondescribed herein will differ from a polypeptide sequence provided herein(e.g., in Table 1) by one or more amino acid substitutions. In someembodiments a sequence will have at least 90% sequence identity (orother degree of sequence identity disclosed hereinbelow) to a sequenceor combination of sequences described herein. In one embodiment thepolypeptide sequence differs from a reference sequence (e.g., inTable 1) by one amino acid substitution. In one embodiment thepolypeptide sequence differs from a reference sequence (e.g., inTable 1) by two amino acid substitutions. In one embodiment thepolypeptide sequence differs from a reference sequence by two amino acidsubstitutions. In one embodiment the polypeptide sequence differs from areference sequence by three amino acid substitutions. In one embodimentthe polypeptide sequence differs from a reference sequence by four aminoacid substitutions. In one embodiment the polypeptide sequence differsfrom a reference sequence by five amino acid substitutions. In oneembodiment the polypeptide sequence differs from a reference sequence bysix amino acid substitutions. In one embodiment the polypeptide sequencediffers from a reference sequence by seven amino acid substitutions. Inone embodiment the polypeptide sequence differs from a referencesequence by eight amino acid substitutions. In one embodiment thepolypeptide sequence differs from a reference sequence by nine aminoacid substitutions. In one embodiment the polypeptide sequence differsfrom a reference sequence by ten amino acid substitutions. In certainembodiments the polypeptide sequence differs from a reference sequenceby 1-10 amino acid substitutions, sometimes 1-5 amino acidsubstitutions. In some cases, amino acid substitutions are selected thatdo not change a basic property (e.g., binding specificity) relative tothe reference sequence. In some cases, amino acid substitutions areselected that change binding affinity but not binding specificity. Insome cases substitutions are selected to change a property (e.g.,substitutions that affect effector function or half-life) as known inthe art or described herein below. In some embodiments the amino acidsubstitutions are conservative substitutions. A conservative amino addsubstitution is recognized in the art as a substitution of one amino addfor another amino add that has similar properties, such as polarity,charge, hydrophobicity, and aromaticity. A conservative amino addsubstitution can also be made based on the side chain characteristics ofthe amino add, such as containing sulfur, hydroxyl, or amide.Non-limiting examples of conservative amino add substitutions are setout below.

Amino acid property Amino acid Polar-uncharged Cys, Ser, Thr, Met, Asn,Gln Polar-charged Asp, Glu, Lys, Arg Non-polar Gly, Ala, Pro, Ile, Leu,Val Aromatic His, Phe, Trp, Tyr Aliphatic Ala, Leu, Ile, Val, ProPositively charged Lys, Arg, His Negatively charged Asp, GluSulfur-containing Met Hydroxyl-containing Ser, Thr, Tyr Amide-containingAsn, Gln Sulfhydryl containing Cys

DETAILED DESCRIPTION OF THE INVENTION I. Introduction

In one aspect, the present disclosure provides human allergen-specificmonoclonal antibodies and methods for generating human allergen-specificmonoclonal antibodies from single IgE- or IgG4-expressing human B cells.IgE antibodies, the least abundant class of antibodies in humans, areknown to cause the symptoms of allergic reactions. For example, foodallergy symptoms ranging from urticaria to potentially fatal anaphylaxisresult from the degranulation of mast cells and basophils induced by therecognition of allergic food proteins by surface-bound IgE antibodies.Despite this central role in immunity and allergic disease, human IgEantibodies remain poorly characterized due to their scarcity.Fitzsimmons et al., Front Immunol., 2014, 5:61. Similarly, there is alack of knowledge, but growing interest, surrounding the IgG4 isotypedue to its potential role in mediating the reduced clinical allergenreactivity that accompanies immunotherapy and early allergen exposurethrough antigen blocking. Tordesillas et al., Immunity, 2017, 47:32-50.

The present disclosure provides therapeutic methods for treating a humansubject having an allergy or reducing one or more allergy symptoms in ahuman subject with one or more of the allergen-specific monoclonalantibodies or antigen-binding portions thereof as disclosed herein. Insome embodiments and without intending to be bound by a particularmechanism, the allergen-specific monoclonal antibodies disclosed hereinare used therapeutically as blocking antibodies, which is often referredto as passive immunotherapy.

As described herein, the methods of the disclosure can be used togenerate, from a sample from a human subject having an allergy to anantigen of interest, a pool of genotype-confirmed IgE or IgG4 single Bcells that are candidates for producing antibodies having high affinityfor an allergen of interest. As described in the Examples section below,it has been found that analyzing the cDNA sequences of immunoglobulinheavy chain constant regions to identify the isotype of single B cellsavoids the problem of isotype mischaracterization that is known to occurwhen B cell isotype is determined based on sorting cells by cell surfacemarkers (e.g., as is typically done in FACS cell surface staining). Thisproblem of isotype mischaracterization is known to be especiallypervasive for IgE B cells because the marker CD23 is a “low-affinity”IgE receptor that captures IgE on the surface of non-IgE B cells. See,Berkowska et al., J Allergy Clin Immunol, 2014, 134:688-697. Thus, themethods of the present disclosure generate a pool of single B cells thatare much more likely to produce antibodies having high affinity for theallergen. Furthermore, it has been found that antibodies generatedaccording to the methods disclosed herein are among the highest affinitynative human antibodies discovered to date and exhibit cross-reactivityto different antigens.

II. Definitions

The terminology used herein is for the purpose of describing particularembodiments only, and is not intended to be limiting, because the scopeof the present invention will be limited only by the appended claims.Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. In this specification and inthe claims that follow, reference will be made to a number of terms thatshall be defined to have the following meanings unless a contraryintention is apparent. In some cases, terms with commonly understoodmeanings are defined herein for clarity and/or for ready reference, andthe inclusion of such definitions herein should not be construed asrepresenting a substantial difference over the definition of the term asgenerally understood in the art.

All numerical designations, e.g., pH, temperature, time, concentration,and molecular weight, including ranges, are approximations which arevaried (+) or (−) by increments of 0.1 or 1.0, as appropriate. It is tobe understood, although not always explicitly stated that all numericaldesignations are preceded by the term “about.”

The singular forms “a,” “an,” and “the” include plural referents unlessthe context clearly dictates otherwise. Thus, for example, reference to“a compound” includes a plurality of compounds.

The term “comprising” is intended to mean that the compounds,compositions and methods include the recited elements, but not excludingothers. “Consisting essentially of” when used to define compounds,compositions and methods, shall mean excluding other elements that wouldmaterially affect the basic and novel characteristics of the claimedinvention. “Consisting of” shall mean excluding any element, step, oringredient not specified in the claim. Embodiments defined by each ofthese transition terms are within the scope of this invention.

The term “allergen” refers to a substance that induces an immuneresponse in a subject that results in an allergic reaction by thesubject.

As used herein, the term “antibody” refers to a polypeptide encoded byan immunoglobulin gene or functional fragments thereof that specificallybinds and recognizes an antigen. The term “antibody,” as used herein,also includes antibody fragments that retain binding specificity,including but not limited to Fab, F(ab′)₂, Fv, and scFv. The recognizedimmunoglobulin genes include the kappa, lambda, alpha, gamma, delta,epsilon, and mu constant region genes, as well as myriad immunoglobulinvariable region genes. Light chains are classified as either kappa orlambda. Heavy chains are classified as gamma, mu, alpha, delta, orepsilon, which in turn define the immunoglobulin classes, IgG, IgM, IgA,IgD and IgE, respectively.

An exemplary immunoglobulin (antibody) structural unit comprises twoidentical pairs of polypeptide chains, each pair having one “light”chain (about 25 kDa) and one “heavy” chain (about 50-70 kDa). TheN-terminus of each chain defines a variable region of about 100 to 110or more amino acids primarily responsible for antigen recognition. Thus,the terms “variable heavy chain” or “VH” refer to the variable region ofan immunoglobulin heavy chain, including an Fv, scFv, dsFv or Fab; whilethe terms “variable light chain” or “VL” refer to the variable region ofan immunoglobulin light chain, including an Fv, scFv, dsFv or Fab.

The term “variable region” refers to a domain in an antibody heavy chainor light chain that gives an antibody its specificity for binding to anantigen. Typically, an antibody variable region comprises four conserved“framework” regions interspersed with three hypervariable“complementarity determining regions.”

The term “complementarity determining region” or “CDR” refers to thethree hypervariable regions in each chain that interrupt the fourframework regions established by the light and heavy chain variableregions. The CDRs are primarily responsible for binding to an epitope ofan antigen. The CDRs of each chain are typically referred to as CDR1,CDR2, and CDR3, numbered sequentially starting from the N-terminus, andare also typically identified by the chain in which the particular CDRis located. Thus, a VH CDR3 is located in the variable domain of theheavy chain of the antibody in which it is found, whereas a VL CDR1 isthe CDR1 from the variable domain of the light chain of the antibody inwhich it is found.

As noted, the part of a variable region not contained in the CDRs iscalled the framework. The “framework regions” of different light orheavy chains are relatively conserved within a species. The frameworkregion of an antibody, that is the combined framework regions of theconstituent light and heavy chains, serves to position and align theCDRs in three-dimensional space. Framework sequences can be obtainedfrom public DNA databases or published references that include germlineantibody gene sequences. For example, germline DNA sequences for humanheavy and light chain variable region genes can be found in the “VBASE2”germline variable gene sequence database for human and mouse sequences.

The amino acid sequences of the CDRs and framework regions can bedetermined using various well known definitions in the art. The positionand length of the CDRs have been precisely defined by Kabat et al.,Sequences of Proteins of Immunological Interest, U.S. Department ofHealth and Human Services, 1983, 1987, and others. See, e.g., Johnsonand Wu, Nucleic Acids Res. 2000 Jan. 1; 28(1): 214-218; Johnson et al.,Nucleic Acids Res., 29:205-206 (2001); Chothia & Lesk, (1987)J. Mol.Biol. 196, 901-917; Chothia et al. (1989) Nature 342, 877-883; Chothiaet al. (1992) J. Mol. Biol. 227, 799-817; Al-Lazikani et al., J. Mol.Biol 1997, 273(4)); and MacCallum et al., J. Mol. Biol., 262:732-745(1996). Also see international ImMunoGeneTics database (IMGT), AbM, andobserved antigen contacts.

The terms “antigen-binding portion” and “antigen-binding fragment” areused interchangeably herein and refer to one or more fragments of anantibody that retains the ability to specifically bind to an antigen(e.g., an allergen, e.g., Ara h 2 or Ara h 3). Examples ofantibody-binding fragments include, but are not limited to, a Fabfragment (a monovalent fragment consisting of the VL, VH, CL, and CH1domains), F(ab′)₂ fragment (a bivalent fragment comprising two Fabfragments linked by a disulfide bridge at the hinge region), a singlechain Fv (scFv), a disulfide-linked Fv (dsFv), complementaritydetermining regions (CDRs), VL (light chain variable region), VH (heavychain variable region), nanobodies, and any combination of those or anyother functional portion of an immunoglobulin peptide capable of bindingto target antigen. Antibodies and antigen-binding portions thereofinclude domain-specific antibodies, single domain antibodies,domain-deleted antibodies, chimeric antibodies, CDR-grafted antibodies,diabodies, triabodies, tetrabodies, minibodies, nanobodies (e.g.monovalent nanobodies, bivalent nanobodies, etc.), small modularimmunopharmaceuticals (SMIPs), and shark variable IgNAR domains.Exemplary configurations of variable and constant domains that may befound within an antigen-binding fragment of an antibody of the presentinvention include: (a) VH-CHI; (b) VH-CH2; (c) VH-CH3; (d) VH-CH1-CH2;(e) VH-Ch1-Ch2-Ch3; (f) VH-Ch2-Ch3; (g) VH-CL; (h) VL-CH1; (i) VL-Ch2;(X) VL-Ch3; (j) VL-CH1-CH2; (k) VL-CH1-CH2-CH3; (l) VL-CH2-CH3; and (m)VL-CL (see, e.g., FUNDAMENTAL IMMUNOLOGY (Paul ed., 4th ed. 2001),Gruber et al. (1994) J Immunol. 152:5368-5374; McCartney, et al., 1995Protein Eng. 8:301-314; Shukra et al., 2014, “Production of recombinantantibodies using bacteriophages” Eur J Microbiol Immunol (Bp). 4(2):91-98; Todorovska, 2001, “Design and application of diabodies,triabodies and tetrabodies for cancer targeting” J Immunol Methods;248(1-2):47-66; Salvador et al., 2019, “Nanobody: outstanding featuresfor diagnostic and therapeutic applications” Anal Bioanal Chem.411(9):1703-1713; Gill et al., 2006, “Biopharmaceutical drug discoveryusing novel protein scaffolds.” Curr Opin Biotechnol., (6):653-8; andUbah et al., 2016, “Phage Display Derived IgNAR V Region Binding Domainsfor Therapeutic Development” Curr Pharm Des. 22(43):6519-6526, each ofwhich is incorporated by reference herein.

The term “epitope” refers to the area or region of an antigen to whichan antibody specifically binds, i.e., an area or region in physicalcontact with the antibody, and can include a few amino acids or portionsof a few amino acids, e.g., 5 or 6, or more, e.g., 20 or more aminoacids, or portions of those amino acids. In some cases, the epitopeincludes non-protein components, e.g., from a carbohydrate, nucleicacid, or lipid. In some cases, the epitope is a three-dimensionalmoiety. Thus, for example, where the target is a protein, the epitopecan be comprised of consecutive amino acids, or amino acids fromdifferent parts of the protein that are brought into proximity byprotein folding (e.g., a discontinuous epitope).

A “monoclonal antibody” refers to antibodies produced by a single cloneof cells or a single cell line and consisting of or consistingessentially of antibody molecules that are identical in their primaryamino acid sequence. In some embodiments, a monoclonal antibodypreparation comprises a population of antibodies that are identical andbind to the same epitope of an antigen, except for mutations that ariseduring monoclonal antibody production. Unless otherwise specified orclear from context, the term ‘monoclonal antibody’ includes syntheticantibodies and antigen binding fragments thereof.

A “human antibody” refers to an antibody having variable and constantregions derived from human germline immunoglobulin sequences. A humanantibody of the present disclosure may include amino acid residues notencoded by human germline immunoglobulin sequences (e.g., mutationsintroduced by random or site-directed mutagenesis in vitro or by somaticmutations in vivo). The term “human antibody” is not intended to includechimeric or humanized antibodies in which CDR sequences derived from thegermline or immune cells of a non-human species (e.g., mouse) have beengrafted onto human framework sequences.

The term “specifically binds” refers to a molecule (e.g., an antibody orantibody fragment) that binds to a target with greater affinity,avidity, more readily, and/or with greater duration to that target in asample than it binds to a non-target compound. In some embodiments, anantibody or antigen-binding portion thereof that specifically binds atarget (e.g., an allergen, e.g., Ara h 2 or Ara h 3) is an antibody orantigen-binding portion that binds to the target with at least 2-foldgreater affinity than non-target compounds, e.g., at least 3-fold,4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 20-fold,25-fold, 50-fold or greater affinity. For example, in some embodiments,an antibody that specifically binds to an allergen target, such as Ara h2 or Ara h 3, will typically bind to the allergen target with at least a2-fold greater affinity than to a non-allergen target. It will beunderstood by a person of ordinary skill in the art that an antibodythat specifically or preferentially binds to a first target may or maynot specifically or preferentially bind to a second target.

The term “binding affinity,” as used herein, refers to the strength of anon-covalent interaction between two molecules, e.g., an antibody (or anantigen-binding fragment thereof) and an antigen. Thus, for example, theterm may refer to 1:1 interactions between an antibody (or anantigen-binding fragment thereof) and an antigen, unless otherwiseindicated or clear from context. Binding affinity may be quantified bymeasuring an equilibrium dissociation constant (K_(D)), which refers tothe dissociation rate constant (k_(d), time⁻¹) divided by theassociation rate constant (k_(a), time⁻¹ M⁻¹). K_(D) can be determinedby measurement of the kinetics of complex formation and dissociation,e.g., using Surface Plasmon Resonance (SPR) methods, e.g., a Biacore™system; kinetic exclusion assays such as KinExA®; and BioLayerinterferometry (e.g., using the ForteBio® Octet platform). As usedherein, “binding affinity” includes not only formal binding affinities,such as those reflecting 1:1 interactions between an antibody (or anantigen-binding fragment thereof) and an antigen, but also apparentaffinities for which Kos are calculated that may reflect avid binding.

The term “cross-reacts,” as used herein, refers to the ability of anantibody to bind to two or more antigens. As a non-limiting example, insome embodiments, an antibody that specifically binds to a firstallergen target (e.g., a first peanut allergen, such as Ara h 2) canexhibit cross-reactivity with a second allergen target (e.g., a secondpeanut allergen, such as Ara h 3).

The term “isolated,” as used with reference to a nucleic acid or protein(e.g., antibody), denotes that the nucleic acid or protein isessentially free of other cellular components with which it isassociated in the natural state. It is preferably in a homogeneousstate. Purity and homogeneity are typically determined using analyticalchemistry techniques such as electrophoresis (e.g., polyacrylamide gelelectrophoresis) or chromatography (e.g., high performance liquidchromatography). In some embodiments, an isolated nucleic acid orprotein (e.g., antibody) is at least 85% pure, at least 90% pure, atleast 95% pure, or at least 99% pure.

The terms “polypeptide,” “peptide,” and “protein” are usedinterchangeably herein to refer to a polymer of amino acid residues. Theterms apply to amino acid polymers in which one or more amino acidresidue is an artificial chemical mimetic of a corresponding naturallyoccurring amino acid, as well as to naturally occurring amino acidpolymers and non-naturally occurring amino acid polymers. As usedherein, the terms encompass amino acid chains of any length, includingfull length proteins, wherein the amino acid residues are linked bycovalent peptide bonds.

The term “amino acid” refers to refers to naturally occurring andsynthetic amino acids, as well as amino acid analogs and amino acidmimetics that function in a manner similar to the naturally occurringamino acids. Naturally occurring amino acids are those encoded by thegenetic code, as well as those amino acids that are later modified,e.g., hydroxyproline, y-carboxyglutamate, and O-phosphoserine. Aminoacid analogs refers to compounds that have the same basic chemicalstructure as a naturally occurring amino acid, i.e., an a carbon that isbound to a hydrogen, a carboxyl group, an amino group, and an R group,e.g., homoserine, norleucine, methionine sulfoxide, methionine methylsulfonium. Such analogs have modified R groups (e.g., norleucine) ormodified peptide backbones, but retain the same basic chemical structureas a naturally occurring amino acid. “Amino acid mimetics” refers tochemical compounds that have a structure that is different from thegeneral chemical structure of an amino acid, but that functions in amanner similar to a naturally occurring amino acid.

Amino acids may be referred to herein by either their commonly knownthree letter symbols or by the one-letter symbols recommended by theIUPAC-IUB Biochemical Nomenclature Commission. Nucleotides, likewise,may be referred to by their commonly accepted single-letter codes.

As used herein, the terms “nucleic acid” and “polynucleotide” are usedinterchangeably. Use of the term “polynucleotide” includesoligonucleotides (i.e., short polynucleotides). This term also refers todeoxyribonucleotides, ribonucleotides, and naturally occurring variants,and can also refer to synthetic and/or non-naturally occurring nucleicacids (i.e., comprising nucleic acid analogues or modified backboneresidues or linkages), such as, for example and without limitation,phosphorothioates, phosphoramidates, methyl phosphonates, chiral-methylphosphonates, 2-O-methyl ribonucleotides, peptide-nucleic acids (PNAs),and the like. Unless otherwise indicated, a particular nucleic acidsequence also implicitly encompasses conservatively modified variantsthereof (e.g., degenerate codon substitutions) and complementarysequences as well as the sequence explicitly indicated. Specifically,degenerate codon substitutions may be achieved by generating sequencesin which the third position of one or more selected (or all) codons issubstituted with mixed-base and/or deoxyinosine residues (see, e.g.,Batzer et al., Nucleic Acid Res. 19:5081 (1991); Ohtsuka et al, J. Biol.Chem. 260:2605-2608 (1985); Rossolini et al, Mol. Cell. Probes 8:91-98(1994)).

The term “sample,” as used herein, refers to a biological sampleobtained from a human or non-human mammalian subject. In someembodiments, a sample comprises blood, blood fractions or blood products(e.g., serum, plasma, platelets, red blood cells, peripheral bloodmononuclear cells and the like); sputum or saliva; stool, urine, otherbiological fluids (e.g., lymph, saliva, prostatic fluid, gastric fluid,intestinal fluid, renal fluid, lung fluid, cerebrospinal fluid, and thelike), tissue (e.g., kidney, lung, liver, heart, brain, nervous tissue,thyroid, eye, skeletal muscle, cartilage, or bone tissue), culturedcells (e.g., primary cultures, explants, transformed cells, or stemcells), or a biopsy sample.

The terms “subject” and “patient,” as used interchangeably herein, referto a mammal, including but not limited to humans, non-human primates,rodents (e.g., rats, mice, and guinea pigs), rabbits, cows, pigs,horses, and other mammalian species. In one embodiment, the subject orpatient is a human.

The terms “treat,” “treating,” and “treatment” refer to any indicia ofsuccess in the treatment or amelioration of an injury, disease, orcondition, including any objective or subjective parameter such asabatement; remission; diminishing of symptoms or making the injury,disease, or condition more tolerable to the subject; slowing in the rateof degeneration or decline; making the final point of degeneration lessdebilitating; and/or improving a subject's physical or mentalwell-being. The treatment or amelioration of symptoms can be based onobjective or subjective parameters. The effect of treatment can becompared to an individual or pool of individuals not receiving thetreatment, or to the same patient prior to treatment or at a differenttime during treatment.

The term “pharmaceutical composition” refers to a composition suitablefor administration to a subject. In general, a pharmaceuticalcomposition is sterile, and preferably free of contaminants that arecapable of eliciting an undesirable response with the subject.Pharmaceutical compositions can be designed for administration tosubjects in need thereof via a number of different routes ofadministration, including oral, intravenous, buccal, rectal, parenteral,intraperitoneal, intradermal, intratracheal, intramuscular,subcutaneous, inhalational, and the like.

The term “pharmaceutically acceptable excipient” refers to a non-activepharmaceutical ingredient that is biologically or pharmacologicallycompatible for use in humans or animals, such as, but not limited to abuffer, carrier, or preservative.

As used herein, a “therapeutic amount” or “therapeutically effectiveamount” of an agent (e.g., a monoclonal antibody as disclosed herein) isan amount of the agent that treats, ameliorates, abates, remits,improves patient survival, increases survival time or rate, diminishessymptoms, makes an injury, disease, or condition (e.g., an allergy) moretolerable, slows the rate of degeneration or decline, or improves apatient's physical or mental well-being. For example, for the givenparameter, a therapeutically effective amount will show an increase ordecrease of therapeutic effect at least 5%, 10%, 15%, 20%, 25%, 40%,50%, 60%, 75%, 80%, 90%, or 100%. Therapeutic efficacy can also beexpressed as “-fold” increase or decrease. For example, atherapeutically effective amount can have at least a 1.2-fold, 1.5-fold,2-fold, 5-fold, or more effect over a control.

The terms “administer,” “administered,” or “administering” refer tomethods of delivering agents, compounds, or compositions to the desiredsite of biological action. These methods include, but are not limitedto, topical delivery, parenteral delivery, intravenous delivery,intradermal delivery, intramuscular delivery, rectal delivery, orintraperitoneal delivery. Administration techniques that are optionallyemployed with the agents and methods described herein, include e.g., asdiscussed in Goodman and Gilman, The Pharmacological Basis ofTherapeutics, current ed.; Pergamon; and Remington's, PharmaceuticalSciences (current edition), Mack Publishing Co., Easton, Pa.

III. Methods of Generating Allergen-Specific Antibodies

In one aspect, methods of generating allergen-specific monoclonalantibodies from a human sample are provided. In some embodiments, themethod comprises:

-   -   (a) isolating single B cells from a sample from a human subject,        wherein the subject is allergic to the allergen;    -   (b) generating cDNAs from the single B cells of step (a),        wherein the cDNA sequences comprise a first sequence that        encodes all or part of an immunoglobulin heavy chain and a        second sequence that encodes all or part of an immunoglobulin        light chain;    -   (c) determining the sequences of the cDNAs from step (b);    -   (d) analyzing the sequences determined in step (c) to identify        single B cells comprising a first sequence that comprises an IgE        constant region or an IgG4 constant region;    -   (e) identifying, from the single B cells of step (d), (i) a        heavy chain variable region sequence in the immunoglobulin heavy        chain that comprises an IgE constant region or an IgG4 constant        region, and (ii) a light chain variable region sequence in an        immunoglobulin light chain that is co-expressed with the        immunoglobulin heavy chain in the same single B cell;    -   (f) expressing antibodies comprising the heavy chain variable        region sequence and the light chain variable region sequence        from step (e); and    -   (g) identifying one or more antibodies from step (f) that        specifically bind to the allergen.

Subject Populations and Samples

In some embodiments, the method of generating allergen-specificmonoclonal antibodies comprises isolating B cells from a biologicalsample from a human subject. In some embodiments, the sample compriseswhole blood, peripheral blood, or a leukapheresis product. In someembodiments, the sample comprises peripheral blood mononuclear cells(PBMCs). In some embodiments, the sample comprises a tissue from thehuman subject, e.g., tonsil tissue, spleen, or bone marrow. Methods ofisolating B cells from blood and tissue samples are described in theart. See, e.g., Heine et al., Curr Protoc. Immunol., 2011,94:7.5.1-7.5.14; and Zuccolo et al., BMC Immunol, 2009, 10:30,doi:10.1186/1471-2172-10-30.

In some embodiments, the allergen-specific antibodies are generated froma human subject having an allergy to the allergen. In some embodiments,the human subject having an allergy is an adult. In some embodiments,the human subject having an allergy is a juvenile. In some embodiments,the human subject has an allergy to a food allergen, a plant allergen, afungal allergen, an animal allergen, a dust mite allergen, a drugallergen, a cosmetic allergen, or a latex allergen. In some embodiments,the human subject has allergies to two or more allergens, e.g., to twoor more of a food allergen, a plant allergen, a fungal allergen, ananimal allergen, a dust mite allergen, a drug allergen, a cosmeticallergen, or a latex allergen. In some embodiments, the human subjecthas allergies to 2, 3, 4, 5, 6, 7, 8, 9, 10 or more allergens. In someembodiments, the human subject has allergies to two or more differenttypes of antigens (allergens) in a class of allergen, e.g., allergies totwo or more different food allergens (e.g., allergies to two or moredifferent peanut antigens, or allergies to a peanut allergen and anon-peanut allergen such as a tree nut, egg, or milk allergen), orallergies to two more different fungal allergens (e.g., allergies to twoor more different species of Aspergillus). In some embodiments, thehuman subject has allergies to two more different classes of allergens(e.g., allergies to one or more food allergens and to one or more plantallergens). In some embodiments, the human subject has allergies to onlyone class of allergens (e.g., the subject has allergies to one or morefood allergens but not to non-food allergens, or the subject hasallergies to one or more fungal allergens but not to non-fungalallergens).

In some embodiments, the human subject has an allergy to a foodallergen. In some embodiments, the food allergen is a milk allergen, anegg allergen, a nut allergen, a tree nut allergen, a fish allergen, ashellfish allergen, a soy allergen, a legume allergen, a seed allergen,or a wheat allergen. In some embodiments, the food allergen is a peanutallergen.

In some embodiments, the human subject has an allergy to a plantallergen or a fungal allergen. In some embodiments, the allergen is afungal allergen (e.g., Aspergillus, e.g., Asp. fumigatus, Asp. niger, orAsp. nidulans). In some embodiments, the allergen is a pollen allergen(e.g., tree pollen, grass pollen, or weed pollen) or a mold allergen. Insome embodiments, the human subject has an allergy to an animalallergen. In some embodiments, the allergen is a dander allergen or aninsect sting.

In some embodiments, the method of generating allergen-specificmonoclonal antibodies does not comprise immunizing the human subjectwith the allergen or exposing the human subject to the allergen prior toobtaining the sample from the subject.

B Cell Isolation and Screening

In some embodiments, single B cells are isolated from the sample fromthe subject having an allergy to the allergen. In some embodiments, thesingle B cells are separated into separate partitions, e.g., separatewells of a multi-well plate, encapsulated into droplets, or dispersedinto microwells. In some embodiments, at least 10, 50, 100, 500, 1,000,5,000, or 10,000 B cells or more are isolated from a sample and areseparated into separate partitions.

In some embodiments, the isolating step comprises sorting cells in thesample by fluorescent activated cell sorting (FACS). FACS sorting can beused to sort cells based on cell surface marker expression, cell size,and/or granularity and deliver cells individually to a well, e.g., a96-well or 384-well tissue culture or PCR plate. Methods of isolatingand purifying cell populations by FACS are described in the art. See,e.g., Basu et al., J Vis Exp, 2010, 41:1546, doi:10.3791/1546.

In some embodiments, a droplet microfluidic platform can be used todispense single B cells into separate droplets. In some embodiments, thenucleic acids (e.g., mRNA) of a single cell in a droplet is labeled witha nucleotide sequence that is unique to the droplet, e.g., a UniqueMolecular Identifier barcode nucleotide sequence, thereby enablingdownstream processing steps for the sequences from multiple B cells tobe performed in a single reaction container. Methods of encapsulatingsingle cells in droplets are described in the art. See, e.g., Macosko etal., Cell, 2015, 161: 1202-1214; Zhang et al., Scientific Reports, 2017,7:41192, doi:10.1038/srep41192.

In some embodiments, cells are dispersed into microwells designed totrap a single cell. Methods of single cell microwell trapping aredescribed in the art. See, e.g. Han et al., Cell, 2018, 172:5,doi:10.1016/j.cell.2018.02.001.

In some embodiments, cells from the sample are screened for thepresence, absence, or level of expression of one or more markers andsingle B cells are isolated based on the presence or level of expressionof the one or more B cell markers (e.g., one, two, three, four, five,six, seven, eight, or more markers). In some embodiments, cells arescreened for the presence, absence, or level of expression of one ormore cell surface B cell markers, such as but not limited to CD19, CD20,CD21, CD22, CD23, CD24, CD40, CD72, or CD79. In some embodiments, a cellis determined to be a B cell if the cell is positive for one or more ofthe B cell markers, e.g., is positive for one or more of CD19, CD20,CD21, CD22, CD23, CD24, CD40, CD72, or CD79. In some embodiments, singleCD19⁺ B cells are isolated.

In some embodiments, cells from the sample are screened for thepresence, absence, or level of expression of one or more immunoglobulinisotypes, such as but not limited to IgE, IgG, IgM, IgA, or IgD or asubclass thereof. In some embodiments, the single B cells that areisolated are selected for expression of cell surface IgE and/or forexpression of cell surface IgG4. In some embodiments, single B cells areisolated without selecting for expression of one or more immunoglobulinisotypes (e.g., without selecting for expression of cell surface IgEand/or for expression of cell surface IgG4).

IgE Selection

In some embodiments, the method comprises isolating single B cells thatare selected for expression of cell surface IgE. In some embodiments,the isolating step comprises contacting cells of the sample with ananti-human IgE antibody and selecting for cells that express IgE on thecell surface. In some embodiments, the isolating step comprisescontacting cells of the sample with antibody against a B cell marker(e.g., an antibody against CD19, CD20, CD21, CD22, CD23, CD24, CD40,CD72, or CD79) and an anti-human IgE antibody and selecting for cellsthat express the B cell marker and that express IgE on the cell surface.In some embodiments, the isolating step comprises contacting cells ofthe sample with an anti-human CD19 antibody and an anti-human IgEantibody and selecting for CD19⁺IgE-expressing B cells.

In some embodiments, the isolating step comprises contacting cells ofthe sample with antibody against a B cell marker (e.g., an antibodyagainst CD19, CD20, CD21, CD22, CD23, CD24, CD40, CD72, or CD79), ananti-human IgE antibody, and an antibody against one or moreimmunoglobulin isotypes (e.g., an anti-human IgG antibody, an anti-humanIgM antibody, an anti-human IgA antibody, and/or an anti-human IgDantibody) or subclass thereof and selecting for cells that express the Bcell marker, that express IgE on the cell surface, and that do notexpress detectable levels of the one or more other immunoglobulinisotypes being screened for. In some embodiments, the isolating stepcomprises contacting cells of the sample with an anti-human CD19antibody, an anti-human IgE antibody, and one or more of an anti-humanIgG antibody, an anti-human IgM antibody, an anti-human IgA antibody, oran anti-human IgD antibody, and selecting for CD19⁺IgE-expressing Bcells that are negative for IgG, IgM, IgA, or IgD cell surfaceexpression.

In some embodiments, the isolating step comprises contacting cells fromthe sample with an anti-human CD19 antibody, an anti-human IgE antibody,an anti-human IgM antibody, and an anti-human IgG antibody and selectingfor CD19⁺IgM⁻IgG⁻ IgE-expressing B cells. In some embodiments, theisolating step comprises contacting cells from the sample with ananti-human CD19 antibody, an anti-human IgE antibody, an anti-human IgMantibody, an anti-human IgG antibody, an anti-human IgA antibody, and ananti-human IgD antibody and selecting for CD19⁺IgM⁻IgG⁻IgA⁻IgD⁻IgE-expressing B cells.

In some embodiments, the isolating step comprises contacting cells ofthe sample with antibody against a B cell marker (e.g., an antibodyagainst CD19, CD20, CD21, CD22, CD23, CD24, CD40, CD72, or CD79) andantibodies against non-IgE isotypes (e.g., an anti-human IgG antibody,an anti-human IgM antibody, an anti-human IgA antibody, and ananti-human IgD antibody) or subclass thereof, and selecting for cellsthat express the B cell marker and that do not express detectable levelsof the non-IgE isotypes. In some embodiments, the isolating stepcomprises contacting cells from the sample with an anti-human CD19antibody, an anti-human IgM antibody, an anti-human IgG antibody, ananti-human IgA antibody, and an anti-human IgD antibody and selectingfor CD19⁺IgM⁻IgG⁻IgA⁻IgD⁻ B cells.

In some embodiments, the method comprises isolating IgE-expressing Bcells that are antibody-secreting B cells (e.g., plasmablasts or plasmacells). In some embodiments, the method comprises isolatingIgE-expressing B cells that are memory B cells. In some embodiments, themethod comprises isolating IgE-expressing antibody-secreting B cells andIgE-expressing memory B cells.

IgG4 Selection

In some embodiments, the method comprises isolating single B cells thatare selected for expression of cell surface IgG4. In some embodiments,the isolating step comprises contacting cells of the sample with ananti-human IgG4 antibody and selecting for cells that express IgG4 onthe cell surface. In some embodiments, the isolating step comprisescontacting cells of the sample with an antibody against a B cell marker(e.g., an antibody against CD19, CD20, CD21, CD22, CD23, CD24, CD40,CD72, or CD79) and an anti-human IgG4 antibody and selecting for cellsthat express the B cell marker and that express IgG4 on the cellsurface. In some embodiments, the isolating step comprises contactingcells of the sample with an anti-human CD19 antibody and an anti-humanIgG4 antibody and selecting for CD19⁺IgG4-expressing B cells.

In some embodiments, the isolating step comprises contacting cells ofthe sample with an antibody against a B cell marker (e.g., an antibodyagainst CD19, CD20, CD21, CD22, CD23, CD24, CD40, CD72, or CD79), ananti-human IgG4 antibody, and an antibody against one or more IgGsubclasses (e.g., an anti-human IgG1 antibody, an anti-human IgG2antibody, an anti-human IgG3 antibody) and selecting for cells thatexpress the B cell marker, that express IgG4 on the cell surface, andthat do not express detectable levels of the one or more other IgGsubclasses being screened for. In some embodiments, the isolating stepcomprises contacting cells from the sample with an anti-human CD19antibody, an anti-human IgG1 antibody, an anti-human IgG2 antibody, ananti-human IgG3 antibody, and an anti-human IgG4 antibody and selectingfor CD19⁺IgG1⁻IgG2⁻IgG3⁻IgG4-expressing B cells.

In some embodiments, the isolating step comprises contacting cells ofthe sample with an antibody against a B cell marker (e.g., an antibodyagainst CD19, CD20, CD21, CD22, CD23, CD24, CD40, CD72, or CD79), ananti-human IgG4 antibody, and an antibody against one or moreimmunoglobulin isotypes (e.g., an anti-human IgE antibody, an anti-humanIgM antibody, an anti-human IgA antibody, and/or an anti-human IgDantibody) or subclass thereof and selecting for cells that express the Bcell marker, that express IgG4 on the cell surface, and that do notexpress detectable levels of the one or more other immunoglobulinisotypes being screened for. In some embodiments, the isolating stepcomprises contacting cells of the sample with an anti-human CD19antibody, an anti-human IgG4 antibody, and one or more of an anti-humanIgE antibody, an anti-human IgM antibody, an anti-human IgA antibody, oran anti-human IgD antibody, and selecting for CD19⁺IgG4-expressing Bcells that are negative for IgE, IgM, IgA, or IgD cell surfaceexpression.

In some embodiments, the isolating step comprises contacting cells fromthe sample with an anti-human CD19 antibody, an anti-human IgG4antibody, an anti-human IgM antibody, and an anti-human IgG antibody andselecting for CD19⁺IgM⁻IgE⁻ IgG4-expressing B cells. In someembodiments, the isolating step comprises contacting cells from thesample with an anti-human CD19 antibody, an anti-human IgG4 antibody, ananti-human IgM antibody, an anti-human IgE antibody, an anti-human IgAantibody, and an anti-human IgD antibody and selecting forCD19⁺IgM⁻IgE⁻IgA⁻IgD⁻ IgG4-expressing B cells.

In some embodiments, the isolating step comprises contacting cells ofthe sample with an antibody against a B cell marker (e.g., an antibodyagainst CD19, CD20, CD21, CD22, CD23, CD24, CD40, CD72, or CD79) andantibodies against non-IgG isotypes (e.g., an anti-human IgE antibody,an anti-human IgM antibody, an anti-human IgA antibody, and ananti-human IgD antibody) or non-IgG4 isotypes thereof, and selecting forcells that express the B cell marker and that do not express detectablelevels of the non-IgG or non-IgG4 isotypes. In some embodiments, theisolating step comprises contacting cells from the sample with ananti-human CD19 antibody, an anti-human IgM antibody, an anti-human IgEantibody, an anti-human IgA antibody, and an anti-human IgD antibody andselecting for CD19⁺IgM⁻IgE⁻IgA⁻IgD⁻ B cells. In some embodiments, theisolating step comprises contacting cells from the sample with ananti-human CD19 antibody, an anti-human IgM antibody, an anti-human IgEantibody, an anti-human IgA antibody, an anti-human IgD antibody, ananti-human IgG1 antibody, an anti-human IgG2 antibody, and an anti-humanIgG3 antibody and selecting for CD19⁺IgM⁻IgE⁻IgA⁻IgD⁻ IgG1⁻IgG2⁻IgG3⁻ Bcells.

In some embodiments, the method comprises isolating IgG4-expressing Bcells that are antibody-secreting B cells. In some embodiments, themethod comprises isolating IgG4-expressing B cells that are memory Bcells. In some embodiments, the method comprises isolatingIgG4-expressing antibody-secreting B cells and IgG4-expressing memory Bcells.

Generating and Sequencing cDNAs

In some embodiments, cDNAs are generated from the isolated single Bcells from the sample (e.g., from single B cells that have been screenedfor expression of an immunoglobulin isotype such as IgE or IgG4, or fromsingle B cells that have not been screened for expression of animmunoglobulin isotype). In some embodiments, cDNA libraries areprepared from the single B cells. In some embodiments, for the cDNAsthat are generated for each single B cell, the cDNA sequences comprise asequence that encodes an immunoglobulin heavy chain and a sequence thatencodes an immunoglobulin light chain.

In some embodiments, cDNAs are generated by reverse transcribing cDNAsequences from RNA (e.g., total RNA or mRNA) from the single B cell andamplifying the cDNA sequences. For generating cDNAs, in someembodiments, the single B cells are lysed and cDNA sequences are reversetranscribed from mRNA present in the cell lysate. In some embodiments,RNA is isolated from the single B cell and cDNAs are reverse transcribedfrom the isolated RNA.

In some embodiments, the method comprises amplifying the transcriptomeof the single B cell. For example, in some embodiments, the methodcomprises reverse transcribing RNA (e.g., polyadenylated mRNA) tosynthesize cDNAs, then amplifying the cDNA, e.g., by PCR. Exemplarymethods for reverse transcribing polyadenylated mRNA and amplifying thetranscriptome of a single cell are described in Darmanis et al., CellReports, 2017, 21:1399-1410, and in Picelli et al., Nature Protocols, 9,2014, 171-181.

In some embodiments, the method comprises amplifying immunoglobulinheavy chain and light chain sequences from the single B cells. Forexample, in some embodiments, the method comprises reverse transcribingRNA (e.g., total RNA) to synthesize cDNAs, then amplifying the cDNAs,e.g., by PCR, using primers for immunoglobulin heavy chain variableregions and constant regions. In some embodiments, the method comprisesreverse transcribing RNA using immunoglobulin-specific primers (e.g.,constant region-specific primers) to synthesize cDNAs comprisingimmunoglobulin sequences, then amplifying the cDNAs using primers forimmunoglobulin heavy chain variable regions and constant regions. Anexemplary method for amplifying immunoglobulin heavy chain and lightchain sequences from a single cell is described in Tiller et al., J.Immunol. Methods, 2008, 329:112-124.

After the cDNAs are generated, in some embodiments, the method comprisesdetermining the sequences of the cDNAs. In some embodiments, the cDNAsare subjected to sequencing. In some embodiments, the method comprisessequencing the transcriptomes of the single B cells. In someembodiments, the method comprises sequencing target genes (e.g.,immunoglobulin genes, e.g., immunoglobulin heavy chain variable regionsand constant regions and immunoglobulin light chain variable regions andconstant regions).

Sequencing methods, including methods for high-throughput sequencing,are known in the art. For example, such sequencing technologies include,but are not limited to, pyrosequencing, sequencing-by-ligation, singlemolecule sequencing, sequence-by-synthesis (SBS), massive parallelclonal, massive parallel single molecule SBS, massive parallel singlemolecule real-time, massive parallel single molecule real-time nanoporetechnology, etc. Morozova and Marra provide a review of some suchtechnologies in Genomics, 92: 255 (2008), herein incorporated byreference in its entirety.

In some embodiments, sequencing comprises high-throughput sequencing. Inhigh-throughput sequencing, parallel sequencing reactions using multipletemplates and multiple primers allows rapid sequencing of genomes orlarge portions of genomes. High throughput sequencing methods includemethods that typically use template amplification and those that do not.Sequencing methods that utilize amplification include pyrosequencingcommercialized by Roche as the 454 technology platforms (e.g., GS 20 andGS FLX), clonal array formation and sequencing by synthesis (SBS)chemistry commercialized by Illumina with systems such as the NextSeq,and the Supported Oligonucleotide Ligation and Detection (SOLiD)platform commercialized by Applied Biosystems. Non-amplificationapproaches, also known as single-molecule sequencing, are exemplified bythe HeliScope platform commercialized by Helicos BioSciences, andplatforms commercialized by VisiGen, Oxford Nanopore Technologies Ltd.,Life Technologies/Ion Torrent, and Pacific Biosciences, respectively.

In some embodiments, an Illumina sequencing platform, such as NextSeq,is used. This sequencing technology utilizes clonal array formation andsequencing by synthesis to produce sequences on a large scale. In thismethod, sequencing templates are immobilized on a flow cell surface,then solid-phase amplification creates copies of each template molecule(up to 1,000 identical copies) in close proximity, forming dense“clusters” of polynucleotide sequences. For sequencing the clusters,fluorescently-labeled nucleotides are used to sequences the clusters onthe flow cell surface in parallel. For each sequencing cycle, a singlelabeled reversible terminator-bound dNTP is added to the nucleic acidchain. The sequence of incorporated nucleotides is determined bydetection of post-incorporation fluorescence, then the fluorescent dyeis removed prior to the next cycle of dNTP addition, resulting inbase-by-base sequencing. Typically sequence read length ranges fromabout 30 nucleotides to over 150 nucleotides. For a target cDNA ofinterest having a longer length, the sequence can be bioinformaticallyreassembled based on overlaps between the short sequencing reads todetermine the sequence of the full-length target cDNA.

Identifying B Cells Having an IgE or IgG4 Isotype

In some embodiments, after the sequences of cDNAs have been determinedfor the single B cells, the method comprises analyzing the sequences ofthe cDNAs to identify single B cells that express an immunoglobulinheavy chain having a constant region that is of the IgE isotype and/orof the IgG4 isotype. As described herein, it has been found thatdetermining the isotype of the B cell based on the sequence of the heavychain transcript, rather than FACS immunoglobulin surface staining,substantially reduces the number of false positive IgE cells in the Bcell population, and thus results in a population of B cells that ismuch more likely to yield antibodies that specifically bind to theallergen to which the human subject who is the source of the B cells isallergic.

In some embodiments, the method comprises identifying a sequenceencoding an immunoglobulin heavy chain that comprises an IgE constantregion. In some embodiments, the method comprises identifying a sequenceencoding an immunoglobulin heavy chain that comprises an IgG4 constantregion. In some embodiments, the cDNA sequence is analyzed by comparingthe sequence to a known IgE constant region sequence or to a known IgG4constant region sequence. For example, a comparison of a cDNA sequenceof interest (e.g., a “test” sequence from a B cell) can be compared to aknown IgE or IgG4 constant region sequence (e.g., a “reference”sequence) by aligning the sequences. Methods of alignment of sequencesfor comparison are known in the art. Optimal alignment of sequences forcomparison can be conducted, e.g., by the local homology algorithm ofSmith & Waterman, Adv. Appl. Math. 2:482 (1981), by the homologyalignment algorithm of Needleman & Wunsch, J. Mol. Biol. 48:443 (1970),by the search for similarity method of Pearson & Lipman, Proc. Nat'l.Acad. Sci. USA 85:2444 (1988), by computerized implementations of thesealgorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin GeneticsSoftware Package, Genetics Computer Group, 575 Science Dr., Madison,Wis.), or by manual alignment and visual inspection. Algorithms that aresuitable for determining percent sequence identity and sequencesimilarity are the BLAST and BLAST 2.0 algorithms, which are describedin Altschul et al. (1990) J. Mol. Biol. 215: 403-410 and Altschul et al.(1977) Nucleic Acids Res. 25: 3389-3402, respectively. Software forperforming BLAST analyses is publicly available through the NationalCenter for Biotechnology Information (NCBI) web site.

For comparing a test sequence to an IgE or IgG4 constant regionreference sequence, in some embodiments, the reference sequence ispublished sequence such as an IgE or IgG4 constant region sequence thatis publicly available in the ImMunoGeneTics (IMGT) database. See, e.g.,Camacho et al., BMC Bioinformatics, 2009, 10:421; Lefranc et al.,Nucleic Acids Res, 2009, 37:D1006-1012. Methods of analyzing testsequences to identify sequences comprising an immunoglobulin heavy chainconstant region that is of the IgE isotype and/or of the IgG4 isotypeare also described in Table 1 below.

In some embodiments, in addition to analyzing sequences to identify andselect single B cells comprising an immunoglobulin heavy chain sequencethat comprises an IgE constant region or an IgG4 constant region, themethod further comprises determining the sequences and/or levels ofexpression of one or more other genes in the single B cell. For example,in some embodiments, the method comprises determining the sequencesand/or levels of expression of a set of genes that are a “signature” fora particular type of B cell.

In some embodiments, for a B cell that is identified as having animmunoglobulin heavy chain that comprises an IgE constant region or anIgG4 constant region, the method further comprises identifying, from thesame B cell, the heavy chain variable region sequence that is expressedby the cell and the light chain variable region sequence that isexpressed by the cell.

Antibody Expression

Typically, for a single B cell that is identified as having a cDNA thatcomprises an IgE or IgG4 constant region sequence, the heavy chainvariable region and light chain variable region sequences from thesingle B cell are candidate antibody sequences for having specificity tothe allergen of interest. Thus, in some embodiments, the methodcomprises expressing antibodies comprising the heavy chain variableregion and light chain variable region sequences from the single B celland identifying whether the expressed antibody specifically binds to theallergen of interest. Methods for the expression and purification ofrecombinant antibodies are described in the art. See, e.g., Frenzel etal., Front Immunol., 2013, 4:217, doi:10.3389/fimmu.2013.00217;Siegemund et al., Methods Mol Biol., 2014, 1131:273-295.

In some embodiments, the heavy chain variable region and light chainvariable region sequences from the single B cell are amplified from thesingle B cell and cloned into an expression vector. In some embodiments,the heavy chain variable region and light chain variable regionsequences from the single B cell are synthesized. In some embodiments,the heavy chain variable region sequence and/or light chain variableregion sequence is codon-optimized, e.g., to increase antibodyexpression by the expression system. See, e.g., Ayyar et al., Methods,2017, 116:51-62.

The heavy chain variable region and light chain variable regionsequences from the single B cell can be expressed using any number ofexpression systems, including prokaryotic and eukaryotic expressionsystems. In some embodiments, the expression system is a mammalian cellexpression, such as a hybridoma, or a CHO or HEK293 cell expressionsystem. Many such systems are widely available from commercialsuppliers. Cell expression systems are also described in the art. See,e.g., Kunert and Reinhart, 2016, “Advances in recombinant antibodymanufacturing” Appl Microbiol Biotechnol. 100:3451-61; Jager et al., BMCProc., 2015, 9:P40, doi:10.1186/1753-6561-9-59-P40; and references citedtherein. In some embodiments, the heavy chain and light chain areexpressed using a single vector, e.g., in a di-cistronic expressionunit, or under the control of different promoters. In other embodiments,the heavy chain and light chain are be expressed using separate vectors.In some embodiments, an expression vector for expressing heavy chainvariable region sequence and/or light chain variable region sequence asdisclosed herein is a vector that comprises a constant region of adesired heavy chain isotype or light chain subclass. For example, aheavy chain variable region sequence as disclosed herein can be clonedinto a vector that comprises a human IgG (e.g., IgG1, IgG2, IgG3, orIgG4) heavy chain constant region, and a light chain variable regionsequence as disclosed herein can be cloned into a vector that comprisesa human lambda or kappa light chain constant region.

After an antibody comprising a heavy chain variable region sequence anda light chain variable region sequence from the single B cell asdisclosed herein is expressed and purified, in some embodiments, themethod comprises determining whether the antibody specifically binds tothe allergen. Methods for analyzing binding affinity and bindingkinetics are known in the art. See, e.g., Ernst et al., Determination ofEquilibrium Dissociation Constants, Therapeutic Monoclonal Antibodies(Wiley & Sons ed. 2009). These methods include, but are not limited to,solid-phase binding assays (e.g., ELISA assay), immunoprecipitation,surface plasmon resonance (SPR, e.g., Biacore™ (GE Healthcare,Piscataway, N.J.)), kinetic exclusion assays (e.g. KinExA®), flowcytometry, fluorescence-activated cell sorting (FACS), BioLayerinterferometry (e.g., Octet™ (FortéBio, Inc., Menlo Park, Calif.)), andWestern blot analysis. SPR techniques are reviewed, e.g., in Hahnfeld etal. Determination of Kinetic Data Using SPR Biosensors, MolecularDiagnosis of Infectious Diseases (2004). In a typical SPR experiment,one interactant (target or targeting agent) is immobilized on anSPR-active, gold-coated glass slide in a flow cell, and a samplecontaining the other interactant is introduced to flow across thesurface. When light of a given wavelength is shined on the surface, thechanges to the optical reflectivity of the gold indicate binding, andthe kinetics of binding. In some embodiments, kinetic exclusion assaysare used to determine affinity. This technique is described, e.g., inDarling et al., Assay and Drug Development Technologies Vol. 2, number 6647-657 (2004). In some embodiments, BioLayer interferometry assays areused to determine affinity. This technique is described, e.g., in Wilsonet al., Biochemistry and Molecular Biology Education, 38:400-407 (2010);Dysinger et al., J. Immunol. Methods, 379:30-41 (2012).

In some embodiments, the expressed antibody specifically binds to theallergen with high affinity. In some embodiments, the antibody has abinding affinity (K_(D)) for the allergen that is less than 250 nM, lessthan 100 nM, less than 50 nM, less than 10 nM, less than 5 nM, less than1 nM, less than 500 pM, less than 250 pM, less than 150 pM, less than100 pM, less than 50 pM, less than 40 pM, less than 30 pM, less than 20pM, less than about 10 pM, or less than 1 pM. In some embodiments, theantibody binds to the allergen with a binding affinity (KD) from 1 nM to250 nM.

Nucleic Acids, Vectors, and Host Cells

In some embodiments, the allergen-specific monoclonal antibodies asdescribed herein are prepared using recombinant methods. Accordingly, insome aspects, the invention provides isolated nucleic acids comprising anucleic acid sequence encoding any of the allergen-specific monoclonalantibodies as described herein (e.g., any one or more of the CDRsdescribed herein); vectors comprising such nucleic acids; and host cellsinto which the nucleic acids are introduced that are used to replicatethe antibody-encoding nucleic acids and/or to express the antibodies. Insome embodiments, the host cell is eukaryotic, e.g., a human cell suchas HEK-293.

In some embodiments, a polynucleotide (e.g., an isolated polynucleotide)comprises a nucleotide sequence encoding an antibody or antigen-bindingportion thereof as described herein (e.g., as described in Section IVbelow). In some embodiments, the polynucleotide comprises a nucleotidesequence encoding one or more amino acid sequences (e.g., CDR, heavychain variable region, or light chain variable region) disclosed inTable 1 below.

In a further aspect, methods of making an allergen-specific monoclonalantibody as described herein are provided. In some embodiments, themethod includes culturing a host cell as described herein (e.g., a hostcell expressing a polynucleotide or vector as described herein) underconditions suitable for expression of the antibody. In some embodiments,the antibody is subsequently recovered from the host cell (or host cellculture medium).

Suitable vectors containing polynucleotides encoding antibodies of thepresent disclosure, or fragments thereof, include cloning vectors andexpression vectors. While the cloning vector selected may vary accordingto the host cell intended to be used, useful cloning vectors generallyhave the ability to self-replicate, may possess a single target for aparticular restriction endonuclease, and/or may carry genes for a markerthat can be used in selecting clones containing the vector. Examplesinclude plasmids and bacterial viruses, e.g., pUC18, pUC19, Bluescript(e.g., pBS SK+) and its derivatives, mpl8, mpl9, pBR322, pMB9, ColE1,pCR1, RP4, phage DNAs, and shuttle vectors such as pSA3 and pAT28.Cloning vectors are available from commercial vendors such as BioRad,Stratagene, and Invitrogen.

Expression vectors generally are replicable polynucleotide constructsthat contain a nucleic acid of the present disclosure. The expressionvector may replicate in the host cells either as episomes or as anintegral part of the chromosomal DNA. Suitable expression vectorsinclude but are not limited to plasmids, viral vectors, includingadenoviruses, adeno-associated viruses, retroviruses, and any othervector.

IV. Monoclonal Antibodies that Specifically Bind to Allergens

In another aspect, allergen-specific monoclonal antibodies, andantigen-binding portions thereof, that are generated from a human sampleaccording to a method disclosed herein are provided. In someembodiments, the monoclonal antibody is an antibody that is generatedaccording to the methods disclosed in Section III above. In someembodiments, the monoclonal antibody is an antibody that is generatedfrom a sample from a human subject having an allergy to a food allergen,a plant allergen, a fungal allergen, an animal allergen, a dust miteallergen, a drug allergen, a cosmetic allergen, or a latex allergen, andthe monoclonal antibody specifically binds to the food allergen, plantallergen, fungal allergen, animal allergen, dust mite allergen, drugallergen, cosmetic allergen, or latex allergen.

In some embodiments, an antibody described herein is a full-lengthantibody, a Fab, a Fab′, a F(ab′)2, a Fab′-SH, an Fv, a single-chainantibody, or a single chain Fv (scFv) antibody. In some embodiments, anantibody described herein comprises an IgG4 constant region. In someembodiments, an antibody described herein is a monospecific antibody. Insome embodiments, an antibody described herein is a multispecificantibody. In particular, an antibody described herein can be abispecific antibody that binds to two different allergens. For example,in some embodiments, an antibody described herein can bind to a peanutallergen and a tree nut allergen. In some embodiments, an antibodydescribed herein can bind to a peanut allergen and a milk allergen. Insome embodiments, an antibody described herein can bind to a peanutallergen and a fungal allergen. In some embodiments, an antibodydescribed herein can bind to a tree nut allergen and a milk allergen. Insome embodiments, an antibody described herein can bind to a tree nutallergen and a fungal allergen. In some embodiments, an antibodydescribed herein can bind to a milk allergen and a fungal allergen.

In some embodiments, the monoclonal antibody or antigen-binding portionthereof is an allergen-specific antibody that comprises a heavy chainvariable region sequence and a light chain variable region sequence thatare identified according to a process comprising:

-   -   (a) isolating single B cells from a sample from a human subject,        wherein the subject is allergic to the allergen;    -   (b) generating cDNAs from the single B cells of step (a),        wherein the cDNA sequences comprise a first sequence that        encodes all or part of an immunoglobulin heavy chain and a        second sequence that encodes all or part of an immunoglobulin        light chain;    -   (c) determining the sequences of the cDNAs from step (b);    -   (d) analyzing the sequences determined in step (c) to identify        single B cells comprising a first sequence that comprises an IgE        constant region or an IgG4 constant region;    -   (e) identifying, from the single B cells of step (d), (i) a        heavy chain variable region sequence in the immunoglobulin heavy        chain that comprises an IgE constant region or an IgG4 constant        region, and (ii) a light chain variable region sequence in an        immunoglobulin light chain that is co-expressed with the        immunoglobulin heavy chain in the same single B cell.

In some embodiments, the heavy chain variable region and the light chainvariable region are from a B cell comprising an immunoglobulin thatcomprises an IgE constant region. In some embodiments, the heavy chainvariable region and the light chain variable region are from a B cellcomprising an immunoglobulin that comprises an IgG4 constant region.

In some embodiments, the monoclonal antibody or antigen-binding portionthereof is an allergen-specific antibody that comprises:

-   -   (a) a heavy chain variable region sequence that is derived from        an immunoglobulin heavy chain from an IgE- or IgG4-producing        single B cell from a human subject who is allergic to the        allergen;    -   (b) a heavy chain IgG constant region sequence;    -   (c) a light chain variable region sequence that is derived from        an immunoglobulin light chain from the IgE- or IgG4-producing        single B cell from a human subject;    -   (d) a light chain constant region sequence that is of the same        class as the immunoglobulin light chain of (c).

In some embodiments, the monoclonal antibody comprises a heavy chainvariable region sequence and a light chain variable region sequence thatare derived from an IgE-producing human B cell. In some embodiments, themonoclonal antibody comprises a heavy chain variable region sequence anda light chain variable region sequence that are derived from anIgG4-producing human B cell.

Characteristics of Allergen-Specific Monoclonal Antibodies

In some embodiments, the monoclonal antibody is an antibody thatspecifically binds to a food allergen, a plant allergen, a fungalallergen, an animal allergen, a dust mite allergen, a drug allergen, acosmetic allergen, or a latex allergen. In some embodiments, themonoclonal antibody is an antibody that specifically binds to a foodallergen, such as a milk allergen, an egg allergen, a nut allergen, afish allergen, a shellfish allergen, a soy allergen, a legume allergen,a seed allergen, or a wheat allergen. In some embodiments, themonoclonal antibody specifically binds to a peanut allergen. In someembodiments, the monoclonal antibody specifically binds to a milkallergen. In some embodiments, the monoclonal antibody specificallybinds to an egg allergen.

In some embodiments, the monoclonal antibody specifically binds to theallergen (e.g., a food allergen, a plant allergen, a fungal allergen, ananimal allergen, a dust mite allergen, a drug allergen, a cosmeticallergen, or a latex allergen) with a binding affinity (K_(D)) of lessthan 100 nM, less than 50 nM, less than 10 nM, less than 5 nM, less than1 nM, less than 500 pM, less than 250 pM, less than 150 pM, less than100 pM, less than 50 pM, less than 40 pM, less than 30 pM, less than 20pM, less than about 10 pM, or less than 1 pM. In some embodiments, theantibody binds to the allergen with a binding affinity (KD) from 1 nM to250 nM.

In some embodiments, the monoclonal antibody exhibits cross-reactivitywith at least two different antigens (e.g., allergens), e.g., at leasttwo food allergens, at least two plant allergens, at least two fungalallergens, at least two animal allergens, at least two dust miteallergens, at least two drug allergens, at least two cosmetic allergens,or at least two latex allergens. In some embodiments, the monoclonalantibody exhibits cross-reactivity with at least two milk allergens, atleast two egg allergens, at least two nut allergens, at least two fishallergens, at least two shellfish allergens, at least two soy allergens,at least two legume allergens, at least two seed allergens, or at leasttwo wheat allergens. It will be appreciated by a person of ordinaryskill in the art that many different allergens, such as many plant foodallergens, can be grouped within a small number of protein families. Forexample, more than half of all plant food allergens can be categorizedinto one of the following four structural protein families: the prolaminsuperfamily, the cupin superfamily, profilins, and Bet v-1-relatedproteins. It will also be appreciated by a person of ordinary skill inthe art that for a particular type of allergen (e.g., a “peanut”allergen), there can be more than one peptide or protein that is anallergen. As a non-limiting example, there are 12 known peanutallergens. See, Mueller et al., Curr Allergy Asthma Rep, 2014, 14:429.In some embodiments, the monoclonal antibody exhibits cross-reactivitywith two or more different antigens that are different types or classesof antigens. As a non-limiting example, in some embodiments, amonoclonal antibody exhibits cross-reactivity with an antigen that is apeanut allergen and an antigen that is a nut (e.g., tree nut) allergen.

In embodiments in which the monoclonal antibody exhibitscross-reactivity with at least two different antigens (e.g., allergens),in some embodiments the monoclonal antibody specifically binds to atleast one of the allergens with a K_(D) of less than 100 nM, less than50 nM, less than 10 nM, less than 5 nM, less than 1 nM, less than 500pM, less than 250 pM, less than 150 pM, less than 100 pM, less than 50pM, less than 40 pM, less than 30 pM, less than 20 pM, less than about10 pM, or less than 1 pM. In some embodiments, the monoclonal antibodyspecifically binds to the first antigen (e.g., first allergen) with aK_(D) of less than 100 nM, less than 50 nM, less than 10 nM, less than 5nM, less than 1 nM, less than 500 pM, less than 250 pM, less than 150pM, less than 100 pM, less than 50 pM, less than 40 pM, less than 30 pM,less than 20 pM, less than about 10 pM, or less than 1 pM. In someembodiments, the monoclonal antibody specifically binds to the secondantigen (e.g., second allergen) with a K_(D) of less than 1 μM, lessthan 500 nM, less than 100 nM, less than 10 nM, or less than 1 nM.

Engineered Variations in Variable Regions

In some embodiments, the heavy chain variable region and/or the lightchain variable region of the monoclonal antibody has an identicalsequence to the heavy chain variable region and/or the light chainvariable region encoded by the IgE-producing or IgG4-producing single Bcell from the human subject having an allergy to the allergen. In someembodiments, the heavy chain variable region and/or the light chainvariable region of the monoclonal antibody comprises one or moremodifications, e.g., amino acid substitutions, deletions, or insertions.

As described in the Examples section below, the heavy chain variableregion sequence and/or light chain variable region sequence of anantibody described herein (e.g., a peanut allergen-specific monoclonalantibody such as Clone PA13P1H08, Clone PA13P1E10, Clone PA12P3F10,Clone PA13P3G09, Clone PA12P3D08, Clone PA12P1C07, Clone PA15P1D12,Clone PA15P1D05, or a Clone PA13P1H08 variant) can be engineered tocomprise one or more variations in the heavy chain variable regionsequence and/or light chain variable region sequence. In someembodiments, the engineered variation(s) improves the binding affinityof the antibody for the allergen. In some embodiments, the engineeredvariation(s) improves the cross-reactivity of the antibody for a secondallergen.

In some embodiments, the engineered variation is a variation in one ormore CDRs, e.g., an amino acid substitution in a heavy chain CDR and/ora light chain CDR as described herein. In some embodiments, theengineered variation is a variation in one or more framework regions,e.g., an amino acid substitution in a heavy chain framework regionand/or a light chain framework region. In some embodiments, theengineered variation is a reversion of a region of the heavy chainand/or light chain sequence to the inferred naïve sequence. Methods fordetermining an inferred naïve immunoglobulin sequence are described inthe art. See, e.g., Magnani et al., PLoS Negl Trop Dis, 2017,11:e0005655, doi:10.1371/journal.pntd.0005655.

In some embodiments, affinity maturation is used to engineer furthermutations that enhance the binding affinity of the antibody for theallergen or enhance the cross-reactivity of the antibody for a secondallergen. Methods for performing affinity maturation are known in theart. See, e.g., Renaut et al., Methods Mol Biol, 2012, 907:451-461.

Constant Regions and Isotype Switching

In some embodiments, the monoclonal antibody comprises a heavy chainvariable region sequence and a light chain variable region sequence thatare derived from an IgE-producing human B cell or from an IgG4-producinghuman B cell, and further comprises a kappa or lambda light chainconstant region. In some embodiments, the light chain constant region(kappa or lambda) is from the same type of light chain (i.e., kappa orlambda) as the light chain variable region that was derived from theIgE-producing human B cell or from an IgG4-producing human B cell; as anon-limiting example, if an IgE-producing human B cell comprises a kappalight chain, then the monoclonal antibody that is produced comprises thelight chain variable region from the IgE-producing B cell and furthercomprises a kappa light chain constant region.

In some embodiments, the monoclonal antibody comprises a heavy chainvariable region sequence and a light chain variable region sequence thatare derived from an IgE-producing human B cell or from an IgG4-producinghuman B cell, and further comprises a heavy chain constant region havingan IgG isotype (e.g., IgG4), an IgA isotype (e.g., IgA1), an IgMisotype, an IgD isotype, or that is derived from an IgG, IgA, IgM, orIgD isotype (e.g., is a modified IgG4 constant region). It will beappreciated by a person of ordinary skill in the art that the differentheavy chain isotypes (IgA, IgD, IgE, IgG, and IgM) have differenteffector functions that are mediated by the heavy chain constant region,and that for certain uses it may be desirable to have an antibody thathas the effector function of a particular isotype (e.g., IgG).

In some embodiments, the monoclonal antibody comprises a native (i.e.,wild-type) human IgG, IgA, IgM, or IgD constant region. In someembodiments, the monoclonal antibody comprises a native human IgG1constant region, a native human IgG2 constant region, a native humanIgG3 constant region, a native human IgG4 constant region, a nativehuman IgA1 constant region, a native human IgA2 constant region, anative human IgM constant region, or a native human IgD constant region.In some embodiments, the monoclonal antibody comprises a heavy chainconstant region that comprises one or more modifications. It will beappreciated by a person of ordinary skill in the art that modificationssuch as amino acid substitutions can be made at one or more residueswithin the heavy chain constant region that modulate effector function.In some embodiments, the modification reduces effector function, e.g.,results in a reduced ability to induce certain biological functions uponbinding to an Fc receptor expressed on an effector cell that mediatesthe effector function. In some embodiments, the modification (e.g.,amino acid substitution) prevents in vivo Fab arm exchange, which canintroduce undesirable effects and reduce the therapeutic efficacy of theantibody. See, e.g., Silva et al., J. Biol Chem, 2015, 280:5462-5469.

In some embodiments, the monoclonal antibody comprises a native (i.e.,wild-type) human IgM constant region, human IgD constant region, humanIgG constant region that is derived from IgG1, IgG2, IgG3, or IgG4, orhuman IgA constant region that is derived from IgA1 or IgA2 andcomprises one or more modifications that modulate effector function. Insome embodiments the monoclonal antibody comprises a human IgM constantregion, human IgD constant region, human IgG constant region that isderived from IgG1, IgG2, IgG3, or IgG4, or human IgA constant regionthat is derived from IgA1 or IgA2. In some embodiments, the monoclonalantibody comprises a native (i.e., wild-type) human IgM constant region,human IgD constant region, human IgG constant region that is derivedfrom IgG1, IgG2, IgG3, or IgG4, or human IgA constant region that isderived from IgA1 or IgA2 and comprises one, two, three, four, five,six, seven, eight, nine, ten or more modifications (e.g., amino acidsubstitutions). In some embodiments the constant regions includesvariations (e.g., one, two, three, four, five, six, seven, eight, nine,ten or more amino acid substitutions) that reduce effector function.

In some embodiments, a monoclonal antibody comprises CDR sequences, aheavy chain variable region, and/or a light chain variable region froman antibody from an IgE or IgG4 B cell as described herein (e.g., asdisclosed in Table 1 below) and further comprises a heavy chain constantregion and/or a light chain constant region that is heterologous to theantibody from the IgE or IgG4 B cell from which the CDR sequences and/orvariable region sequences are derived. For example, in some embodiments,the monoclonal antibody comprises the CDR sequences and/or variableregion sequences of an antibody from an IgE B cell, and furthercomprises a heavy chain constant region and a light chain constantregion that is heterologous to the antibody from the IgE B cell (e.g.,the heavy chain constant region and/or light chain constant region is awild-type or modified IgG1, IgG2, IgG3, or IgG4 constant region, or theheavy chain constant region and/or light chain constant region comprisesone or more modifications (e.g., amino acid substitutions) relative tothe native constant region of the antibody from the IgE B cell).

Antibodies that Specifically Bind to Peanut and/or Tree Nut Allergens

In some embodiments, a monoclonal antibody or antigen-binding portionthereof as disclosed herein specifically binds to a peanut allergenand/or a tree nut allergen. In some embodiments, the monoclonal antibodyspecifically binds to a peanut allergen. In some embodiments, themonoclonal antibody specifically binds to a peanut allergen that is Arah 1, Ara h 2, Ara h 3, or Ara h 6.

In some embodiments, the monoclonal antibody exhibits cross-reactivitywith at least two peanut allergens. In some embodiments, the monoclonalantibody exhibits cross-reactivity with two or more of the peanutallergens Ara h 1, Ara h 2, Ara h 3, and Ara h 6. In some embodiments,the monoclonal antibody specifically binds to at least one of the peanutallergens with a K_(D) of less than 100 nM, less than 50 nM, less than10 nM, less than 5 nM, less than 1 nM, less than 500 pM, less than 250pM, less than 150 pM, less than 100 pM, less than 50 pM, less than 40pM, less than 30 pM, less than 20 pM, less than about 10 pM, or lessthan 1 pM. In some embodiments, the monoclonal antibody specificallybinds to a first peanut allergen with a K_(D) of less than 100 nM (e.g.,less than 50 nM, less than 10 nM, less than 5 nM, or less than 1 nM) andspecifically binds to a second peanut allergen with a K_(D) of less than1 μM (e.g., less than 500 nM, less than 100 nM, less than 10 nM, or lessthan 1 nM). In some embodiments, the monoclonal antibody specificallybinds to Ara h 2 with a K_(D) of less than 100 nM (e.g., less than 50nM, less than 10 nM, less than 5 nM, or less than 1 nM) and specificallybinds to Ara h 1, Ara h 3, or Ara h 6 with a K_(D) of less than 1 μM(e.g., less than 500 nM, less than 100 nM, less than 10 nM, or less than1 nM).

In some embodiments, the monoclonal antibody recognizes an epitope thatcomprises or consists of the amino acid motif DPYSPS (SEQ ID NO:704). Insome embodiments, the monoclonal antibody recognizes an epitope thatcomprises or consists of the amino acid sequence DSYGRDPYSPS (SEQ IDNO:705), YSPSQDPYSPS (SEQ ID NO:706), or PDRRDPYSPS (SEQ ID NO:707).

In some embodiments, the monoclonal antibody or antigen-binding portionthereof specifically binds to a tree nut allergen. In some embodiments,the tree nut allergen is a cashew, pistachio, almond, pine nut, pecan,walnut, hazelnut, or macadamia nut allergen. In some embodiments, themonoclonal antibody exhibits cross-reactivity with at least two tree nutallergens. In some embodiments, the monoclonal antibody exhibitscross-reactivity with both cashew and pistachio allergens. In someembodiments, the monoclonal antibody exhibits cross-reactivity with bothpecan and walnut allergens. In some embodiments, the monoclonal antibodyexhibits cross-reactivity with two or more of pecan, walnut, hazelnut,and macadamia nut allergens. In some embodiments, the monoclonalantibody specifically binds to at least one of the tree nut allergenswith a K_(D) of less than 100 nM, less than 50 nM, less than 10 nM, lessthan 5 nM, less than 1 nM, less than 500 pM, less than 250 pM, less than150 pM, less than 100 pM, less than 50 pM, less than 40 pM, less than 30pM, less than 20 pM, less than about 10 pM, or less than 1 pM.

In some embodiments, the monoclonal antibody or antigen-binding portionthereof specifically binds to a peanut allergen and to a tree nutallergen. In some embodiments, the monoclonal antibody specificallybinds to a peanut allergen and to one or more (e.g., 1, 2, 3, 4, ormore) of a cashew, pistachio, almond, pine nut, pecan, walnut, hazelnut,or macadamia nut allergen.

Peanut-Specific Antibody Sequences

In some embodiments, a monoclonal antibody that specifically binds to apeanut allergen (e.g., that specifically binds to Ara h 1, Ara h 2, Arah 3, or Ara h 6) comprises heavy chain CDRs and/or light chain CDRs thatare disclosed in Table 1 below. In some embodiments, a monoclonalantibody that specifically binds to a peanut allergen comprises a lightchain variable region sequence and/or a heavy chain variable regionsequence that is disclosed in Table 1 below. In some embodiments, amonoclonal antibody that specifically binds to a peanut allergencomprises: a heavy chain variable region comprising an amino acidsequence that has at least 75% sequence identity (e.g., at least 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequenceidentity to a heavy chain variable region sequence disclosed in Table 1and comprises a CDR1, a CDR2, and a CDR3 that is identical to the CDRsof that heavy chain variable region sequence, and a light chain variableregion comprising an amino acid sequence that has at least 70% sequenceidentity (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%,96%, 97%, 98%, or 99% sequence identity to a light chain variable regionsequence disclosed in Table 1 and comprises a CDR1, a CDR2, and a CDR3that is identical to the CDRs of that light chain variable regionsequence.

In some embodiments, a monoclonal antibody that specifically binds to apeanut allergen (e.g., that specifically binds to Ara h 1, Ara h 2, Arah 3, or Ara h 6) comprises a light chain sequence, or a portion thereof,and/or a heavy chain sequence, or a portion thereof, derived from any ofthe following antibodies described herein: Clone PA13P1H08, ClonePA13P1E10, Clone PA12P3F10, Clone PA13P3G09, Clone PA12P3D08, ClonePA12P1C07, Clone PA15P1D12, Clone PA15P1D05, a variant of ClonePA13P1H08 (e.g., an R-R variant, an R-N variant, an N-R variant, anrCDR1-N variant, an rCDR2-N variant, an rCDR3-N variant, or an rFWRs-Nvariant of Clone PA13P1H08), Clone PA12P4D02, Clone PA12P3E09, ClonePA12P3E11, Clone PA12P1D02, Clone PA12P1G11, Clone PA13P1H03, ClonePA12P3C01, or Clone PA12P3E04. The amino acid sequences of the CDR,light chain variable domain (VL), and heavy chain variable domain (VH)of Clone PA13P1H08, Clone PA13P1E10, Clone PA12P3F10, Clone PA13P3G09,Clone PA12P3D08, Clone PA12P1C07, Clone PA15P1D12, Clone PA15P1D05,Clone PA13P1H08 variants, Clone PA12P4D02, Clone PA12P3E09, ClonePA12P3E11, Clone PA12P1D02, Clone PA12P1G11, Clone PA13P1H03, ClonePA12P3C01, and Clone PA12P3E04 are set forth in Table 1 below.

In some embodiments, a monoclonal antibody that specifically binds to apeanut allergen comprises one or more (e.g., one, two, three, four,five, or all six) of:

-   -   (a) a heavy chain CDR1 comprising the amino acid sequence of any        one of SEQ ID NOs:2, 10, 17, 25, 33, 41, 47, 58, 113, 129, 199,        341, 348, 409, 459, or 593;    -   (b) a heavy chain CDR2 comprising the amino acid sequence of any        one of SEQ ID NOs:3, 11, 18, 26, 34, 48, 59, 130, 200, 342, 349,        410, 460, 539, or 594;    -   (c) a heavy chain CDR3 comprising the amino acid sequence of any        one of SEQ ID NOs:4, 12, 19, 27, 35, 42, 49, 55, 60, 131, 201,        350, 411, 461, 540, or 595;    -   (d) a light chain CDR1 comprising the amino acid sequence of any        one of SEQ ID NOs:6, 14, 21, 29, 37, 44, 51, 62, 133, 203, 343,        352, 413, 463, 542, or 597;    -   (e) a light chain CDR2 comprising the amino acid sequence of any        one of SEQ ID NOs:7, 15, 22, 30, 38, 52, 78, 86, 126, 149, 196,        345, 353, or 598; and    -   (f) a light chain CDR3 comprising the amino acid sequence of any        one of SEQ ID NOs:8, 23, 31, 39, 45, 53, 63, 134, 204, 346, 354,        414, 464, 543, or 599.

In some embodiments, a monoclonal antibody that specifically binds to apeanut allergen comprises a heavy chain CDR1-3 and a light chain CDR1-3comprising the amino acid sequences of:

-   -   (a) SEQ ID NOs: 2, 3, 4, 6, 7, and 8, respectively; or    -   (b) SEQ ID NOs: 10, 11, 12, 14, 15, and 8, respectively; or    -   (c) SEQ ID NOs: 17, 18, 19, 21, 22, and 23, respectively; or    -   (d) SEQ ID NOs: 25, 26, 27, 29, 30, and 31, respectively; or    -   (e) SEQ ID NOs: 33, 34, 35, 37, 38, and 39, respectively; or    -   (f) SEQ ID NOs: 41, 34, 35, 37, 38, and 39, respectively; or    -   (g) SEQ ID NOs: 47, 48, 49, 51, 52, and 53, respectively; or    -   (h) SEQ ID NOs: 47, 48, 55, 51, 52, and 53, respectively; or    -   (i) SEQ ID NOs: 58, 59, 60, 62, 30, and 63, respectively; or    -   (j) SEQ ID NOs: 58, 59, 60, 6, 7, and 8, respectively; or    -   (k) SEQ ID NOs: 2, 3, 4, 62, 30, and 63, respectively; or    -   (l) SEQ ID NOs: 58, 3, 4, 6, 7, and 8, respectively; or    -   (m) SEQ ID NOs: 2, 59, 4, 6, 7, and 8, respectively; or    -   (n) SEQ ID NOs: 2, 3, 60, 6, 7, and 8, respectively; or    -   (o) SEQ ID NOs:129, 130, 131, 133, 126, and 134, respectively;        or    -   (p) SEQ ID NOs:341, 342, 343, 345, 78, and 346, respectively; or    -   (q) SEQ ID NOs:348, 349, 350, 352, 353, and 354, respectively;        or    -   (r) SEQ ID NOs:199, 200, 201, 203, 149, and 204, respectively;        or    -   (s) SEQ ID NOs:409, 410, 411, 413, 86, and 414, respectively; or    -   (t) SEQ ID NOs:459, 460, 461, 463, 196, and 464, respectively;        or    -   (u) SEQ ID NOs:113, 539, 540, 542, 196, and 543, respectively;        or    -   (v) SEQ ID NOs:593, 594, 595, 597, 598, and 599, respectively.

In some embodiments, a monoclonal antibody that specifically binds to apeanut allergen comprises a heavy chain variable region comprising anamino acid sequence that has at least 75% sequence identity (e.g., atleast 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to any one of SEQ ID NOs:1, 9, 16, 24, 32, 40, 46,54, 57, 64, 65, 66, 67, 128, 340, 347, 406, 408, 458, 538, or 592. Insome embodiments, a monoclonal antibody that specifically binds to apeanut allergen comprises a light chain variable region comprising anamino acid sequence that has at least 75% sequence identity (e.g., atleast 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to any one of SEQ ID NOs:5, 13, 20, 28, 36, 43, 50,56, 61, 132, 344, 351, 407, 412, 462, 541, or 596. In some embodiments,a monoclonal antibody that specifically binds to a peanut allergencomprises a heavy chain variable region comprising an amino acidsequence that has at least 75% sequence identity (e.g., at least 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequenceidentity) to any one of SEQ ID NOs:1, 9, 16, 24, 32, 40, 46, 54, 57, 64,65, 66, 67, 128, 340, 347, 406, 408, 458, 538, or 592, and comprises alight chain variable region comprising an amino acid sequence that hasat least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%,93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to any one ofSEQ ID NOs: 5, 13, 20, 28, 36, 43, 50, 56, 61, 132, 344, 351, 407, 412,462, 541, or 596.

In some embodiments, a monoclonal antibody that specifically binds to apeanut allergen comprises a heavy chain variable region comprising anamino acid sequence that has at least 75% sequence identity (e.g., atleast 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to any one of SEQ ID NOs: 1, 9, 16, 24, 32, 40, 46,54, 57, 64, 65, 66, 67, 128, 340, 347, 406, 408, 458, 538, or 592 andcomprises a CDR1, a CDR2, and a CDR3 that is identical to the CDRs ofthat SEQ ID NO. In some embodiments, a monoclonal antibody thatspecifically binds to a peanut allergen comprises a light chain variableregion comprising an amino acid sequence that has at least 70% sequenceidentity (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%,96%, 97%, 98%, or 99% sequence identity) to any one of SEQ ID NOs: 5,13, 20, 28, 36, 43, 50, 56, 61, 132, 344, 351, 407, 412, 462, 541, or596 and comprises a CDR1, a CDR2, and a CDR3 that is identical to theCDRs of that SEQ ID NO. In some embodiments, a monoclonal antibody thatspecifically binds to a peanut allergen comprises:

-   -   (a) a heavy chain variable region comprising an amino acid        sequence that has at least 75% sequence identity (e.g., at least        80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%        sequence identity) to any one of SEQ ID NOs: 1, 9, 16, 24, 32,        40, 46, 54, 57, 64, 65, 66, 67, 128, 340, 347, 406, 408, 458,        538, or 592 and that comprises a CDR1, a CDR2, and a CDR3 that        is identical to the CDRs of that SEQ ID NO; and    -   (b) a light chain variable region comprising an amino acid        sequence that has at least 75% sequence identity (e.g., at least        80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%        sequence identity) to any one of SEQ ID NOs: 5, 13, 20, 28, 36,        43, 50, 56, 61, 132, 344, 351, 407, 412, 462, 541, or 596 and        that comprises a CDR1, a CDR2, and a CDR3 that is identical to        the CDRs of that SEQ ID NO.

Clone PA13P1H08

In some embodiments, a monoclonal antibody that specifically binds to apeanut allergen comprises a heavy chain CDR1 comprising the amino acidsequence of SEQ ID NO:2, a heavy chain CDR2 comprising the amino acidsequence of SEQ ID NO:3, a heavy chain CDR3 comprising the amino acidsequence of SEQ ID NO:4, a light chain CDR1 comprising the amino acidsequence of SEQ ID NO:6, a light chain CDR2 comprising the amino acidsequence of SEQ ID NO:7, and a light chain CDR3 comprising the aminoacid sequence of SEQ ID NO:8.

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:1, and that comprises a        heavy chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:2, 3, and 4, respectively; and    -   (b) a light chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:5, and that comprises the        light chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:6, 7, and 8, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:1, and comprises a light chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:5. In some embodiments, the antibodycomprises a heavy chain variable region comprising the amino acidsequence of SEQ ID NO:1 and comprises a light chain variable regioncomprising the amino acid sequence of SEQ ID NO:5.

In some embodiments, the antibody is an antibody that competes forbinding with an antibody as described herein (e.g., an antibodycomprising a heavy chain CDR1-3 and a light chain CDR1-3 comprising theamino acid sequences of SEQ ID NOs:2, 3, 4, 6, 7, and 8, respectively,or an antibody comprising a heavy chain variable region comprising theamino acid sequence of SEQ ID NO:1 and further comprising a light chainvariable region comprising the amino acid sequence of SEQ ID NO:5).

Clone PA13P1E10

In some embodiments, a monoclonal antibody that specifically binds to apeanut allergen comprises a heavy chain CDR1 comprising the amino acidsequence of SEQ ID NO:10, a heavy chain CDR2 comprising the amino acidsequence of SEQ ID NO:11, a heavy chain CDR3 comprising the amino acidsequence of SEQ ID NO:12, a light chain CDR1 comprising the amino acidsequence of SEQ ID NO:14, a light chain CDR2 comprising the amino acidsequence of SEQ ID NO:15, and a light chain CDR3 comprising the aminoacid sequence of SEQ ID NO:8.

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:9, and that comprises a        heavy chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:10, 11, and 12, respectively; and    -   (b) a light chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:13, and that comprises        the light chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:14, 15, and 8, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:9, and comprises a light chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:13. In some embodiments, the antibodycomprises a heavy chain variable region comprising the amino acidsequence of SEQ ID NO:9 and comprises a light chain variable regioncomprising the amino acid sequence of SEQ ID NO:13.

In some embodiments, the antibody is an antibody that competes forbinding with an antibody as described herein (e.g., an antibodycomprising a heavy chain CDR1-3 and a light chain CDR1-3 comprising theamino acid sequences of SEQ ID NOs:10, 11, 12, 14, 15, and 8respectively, or an antibody comprising a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO:9 and further comprisinga light chain variable region comprising the amino acid sequence of SEQID NO:13).

Clone PA12P3F10

In some embodiments, a monoclonal antibody that specifically binds to apeanut allergen comprises a heavy chain CDR1 comprising the amino acidsequence of SEQ ID NO:17, a heavy chain CDR2 comprising the amino acidsequence of SEQ ID NO:18, a heavy chain CDR3 comprising the amino acidsequence of SEQ ID NO:19, a light chain CDR1 comprising the amino acidsequence of SEQ ID NO:21, a light chain CDR2 comprising the amino acidsequence of SEQ ID NO:22, and a light chain CDR3 comprising the aminoacid sequence of SEQ ID NO:23.

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:16, and that comprises a        heavy chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:17, 18, and 19, respectively; and    -   (b) a light chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:20, and that comprises        the light chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:21, 22, and 23, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:16, and comprises a light chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:20. In some embodiments, the antibodycomprises a heavy chain variable region comprising the amino acidsequence of SEQ ID NO:16 and comprises a light chain variable regioncomprising the amino acid sequence of SEQ ID NO:20.

In some embodiments, the antibody is an antibody that competes forbinding with an antibody as described herein (e.g., an antibodycomprising a heavy chain CDR1-3 and a light chain CDR1-3 comprising theamino acid sequences of SEQ ID NOs:17, 18, 19, 21, 22, and 23,respectively, or an antibody comprising a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO:16 and furthercomprising a light chain variable region comprising the amino acidsequence of SEQ ID NO:20).

Clone PA13P3G09

In some embodiments, a monoclonal antibody that specifically binds to apeanut allergen comprises a heavy chain CDR1 comprising the amino acidsequence of SEQ ID NO:25, a heavy chain CDR2 comprising the amino acidsequence of SEQ ID NO:26, a heavy chain CDR3 comprising the amino acidsequence of SEQ ID NO:27, a light chain CDR1 comprising the amino acidsequence of SEQ ID NO:29, a light chain CDR2 comprising the amino acidsequence of SEQ ID NO:30, and a light chain CDR3 comprising the aminoacid sequence of SEQ ID NO:31.

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:24, and that comprises a        heavy chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:25, 26, and 27, respectively; and    -   (b) a light chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:28, and that comprises        the light chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:29, 30, and 31, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:24, and comprises a light chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:28. In some embodiments, the antibodycomprises a heavy chain variable region comprising the amino acidsequence of SEQ ID NO:24 and comprises a light chain variable regioncomprising the amino acid sequence of SEQ ID NO:28.

In some embodiments, the antibody is an antibody that competes forbinding with an antibody as described herein (e.g., an antibodycomprising a heavy chain CDR1-3 and a light chain CDR1-3 comprising theamino acid sequences of SEQ ID NOs:25, 26, 27, 29, 30, and 31,respectively, or an antibody comprising a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO:24 and furthercomprising a light chain variable region comprising the amino acidsequence of SEQ ID NO:28).

Clone PA12P3D08

In some embodiments, a monoclonal antibody that specifically binds to apeanut allergen comprises a heavy chain CDR1 comprising the amino acidsequence of SEQ ID NO:33, a heavy chain CDR2 comprising the amino acidsequence of SEQ ID NO:34, a heavy chain CDR3 comprising the amino acidsequence of SEQ ID NO:35, a light chain CDR1 comprising the amino acidsequence of SEQ ID NO:37, a light chain CDR2 comprising the amino acidsequence of SEQ ID NO:38, and a light chain CDR3 comprising the aminoacid sequence of SEQ ID NO:39.

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:32, and that comprises a        heavy chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:33, 34, and 35, respectively; and    -   (b) a light chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:36, and that comprises        the light chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:37, 38, and 39, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:32, and comprises a light chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:36. In some embodiments, the antibodycomprises a heavy chain variable region comprising the amino acidsequence of SEQ ID NO:32 and comprises a light chain variable regioncomprising the amino acid sequence of SEQ ID NO:36.

In some embodiments, the antibody is an antibody that competes forbinding with an antibody as described herein (e.g., an antibodycomprising a heavy chain CDR1-3 and a light chain CDR1-3 comprising theamino acid sequences of SEQ ID NOs:33, 34, 35, 37, 38, and 39,respectively, or an antibody comprising a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO:32 and furthercomprising a light chain variable region comprising the amino acidsequence of SEQ ID NO:36).

Clone PA12P1C07

In some embodiments, a monoclonal antibody that specifically binds to apeanut allergen comprises a heavy chain CDR1 comprising the amino acidsequence of SEQ ID NO:41, a heavy chain CDR2 comprising the amino acidsequence of SEQ ID NO:34, a heavy chain CDR3 comprising the amino acidsequence of SEQ ID NO:42, a light chain CDR1 comprising the amino acidsequence of SEQ ID NO:44, a light chain CDR2 comprising the amino acidsequence of SEQ ID NO:30, and a light chain CDR3 comprising the aminoacid sequence of SEQ ID NO:45.

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:40, and that comprises a        heavy chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:41, 34, and 42, respectively; and    -   (b) a light chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:43, and that comprises        the light chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:44, 30, and 45, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:40, and comprises a light chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:43. In some embodiments, the antibodycomprises a heavy chain variable region comprising the amino acidsequence of SEQ ID NO:40 and comprises a light chain variable regioncomprising the amino acid sequence of SEQ ID NO:43.

In some embodiments, the antibody is an antibody that competes forbinding with an antibody as described herein (e.g., an antibodycomprising a heavy chain CDR1-3 and a light chain CDR1-3 comprising theamino acid sequences of SEQ ID NOs:41, 34, 42, 44, 30, and 45,respectively, or an antibody comprising a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO:40 and furthercomprising a light chain variable region comprising the amino acidsequence of SEQ ID NO:43).

Clone PA15P1D12

In some embodiments, a monoclonal antibody that specifically binds to apeanut allergen comprises a heavy chain CDR1 comprising the amino acidsequence of SEQ ID NO:47, a heavy chain CDR2 comprising the amino acidsequence of SEQ ID NO:48, a heavy chain CDR3 comprising the amino acidsequence of SEQ ID NO:49, a light chain CDR1 comprising the amino acidsequence of SEQ ID NO:51, a light chain CDR2 comprising the amino acidsequence of SEQ ID NO:52, and a light chain CDR3 comprising the aminoacid sequence of SEQ ID NO:53.

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:46, and that comprises a        heavy chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:47, 48, and 49, respectively; and    -   (b) a light chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:50, and that comprises        the light chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:51, 52, and 53, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:46, and comprises a light chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:50. In some embodiments, the antibodycomprises a heavy chain variable region comprising the amino acidsequence of SEQ ID NO:46 and comprises a light chain variable regioncomprising the amino acid sequence of SEQ ID NO:50.

In some embodiments, the antibody is an antibody that competes forbinding with an antibody as described herein (e.g., an antibodycomprising a heavy chain CDR1-3 and a light chain CDR1-3 comprising theamino acid sequences of SEQ ID NOs:41, 34, 42, 44, 30, and 45,respectively, or an antibody comprising a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO:46 and furthercomprising a light chain variable region comprising the amino acidsequence of SEQ ID NO:50).

Clone PA15P1D05

In some embodiments, a monoclonal antibody that specifically binds to apeanut allergen comprises a heavy chain CDR1 comprising the amino acidsequence of SEQ ID NO:47, a heavy chain CDR2 comprising the amino acidsequence of SEQ ID NO:48, a heavy chain CDR3 comprising the amino acidsequence of SEQ ID NO:55, a light chain CDR1 comprising the amino acidsequence of SEQ ID NO:51, a light chain CDR2 comprising the amino acidsequence of SEQ ID NO:52, and a light chain CDR3 comprising the aminoacid sequence of SEQ ID NO:53.

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:54, and that comprises a        heavy chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:47, 48, and 55, respectively; and    -   (b) a light chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:56, and that comprises        the light chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:51, 52, and 53, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:54, and comprises a light chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:56. In some embodiments, the antibodycomprises a heavy chain variable region comprising the amino acidsequence of SEQ ID NO:54 and comprises a light chain variable regioncomprising the amino acid sequence of SEQ ID NO:56.

In some embodiments, the antibody is an antibody that competes forbinding with an antibody as described herein (e.g., an antibodycomprising a heavy chain CDR1-3 and a light chain CDR1-3 comprising theamino acid sequences of SEQ ID NOs:47, 48, 55, 51, 52, and 53,respectively, or an antibody comprising a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO:54 and furthercomprising a light chain variable region comprising the amino acidsequence of SEQ ID NO:56).

Variant Sequences

In some embodiments, the allergen-specific monoclonal antibody comprisesone or more variations (e.g., amino acid substitutions) in one or moreCDR, heavy chain, and/or light chain sequences as disclosed herein(e.g., one or more mutations in one or more CDR, heavy chain, and/orlight chain sequences of Clone PA13P1H08, Clone PA13P1E10, ClonePA12P3F10, Clone PA13P3G09, Clone PA12P3D08, Clone PA12P1C07, ClonePA15P1D12, Clone PA15P1D05). In some embodiments, one or moresubstitutions are made in a CDR, heavy chain, or light chain sequence ofClone PA13P1H08. As described in the Examples section below, variantswere made of Clone PA13P1H08, in which one or more regions in the heavychain and/or light chain were reverted to the inferred naïverearrangement. It was found that antibody sequences comprising a naïvelight chain or heavy chain sequence were capable of binding to thepeanut allergen Ara h 2. It was also surprisingly found that a variantof Clone PA13P1H08 comprising a reverted CDR-H2 sequence exhibitedsignificantly improved cross-reactivity to a second peanut allergen (Arah 3) in addition to having sub-nanomolar affinity for the Ara h 2 peanutallergen. Thus, in some embodiments, the mutation is an amino acidsubstitution that reverts at least a portion of the sequence of theclone from its “native” form (i.e., the CDR, heavy chain variableregion, or light chain variable region sequence of the clone asdisclosed in Table 1) to the inferred naïve immunoglobulin sequence.

In some embodiments, an allergen-specific monoclonal antibody comprisesone or more variant sequences of a Clone PA13P1H08 variant as disclosedherein. In some embodiments, the antibody comprises one of the sequencesof Clone PA13P1H08 variant “R-R,” in which both the heavy chain variableregion and the light chain variable region of Clone PA13P1H08 arereverted back to the inferred naïve rearrangement. In some embodiments,the antibody comprises a reverted heavy chain variable region sequencecomprising SEQ ID NO:57. In some embodiments, the antibody comprises areverted light chain variable region sequence comprising SEQ ID NO:61.

In some embodiments, the antibody comprises one of the sequences ofClone PA13P1H08 variant “R-N,” in which the heavy chain variable regionof Clone PA13P1H08 is reverted back to the inferred naïve rearrangementand the light chain variable region retains the native sequence of ClonePA13P1H08 (i.e., SEQ ID NO:5). In some embodiments, the antibodycomprises one of the sequences of Clone PA13P1H08 variant “N-R,” inwhich the heavy chain variable region retains the native sequence ofClone PA13P1H08 (i.e., SEQ ID NO:1), and the light chain variable regionis reverted back to the inferred naïve rearrangement of

Clone PA13P1H08.

In some embodiments, the antibody comprises one or more reverted CDRsequences, e.g., one or more reverted heavy chain CDR sequences, and/orone or more reverted light chain CDR sequences. In some embodiments, theantibody comprises one or more of a reverted CDR-H1 comprising SEQ IDNO:58, a reverted CDR-H2 comprising SEQ ID NO:59, or a reverted CDR-H3comprising SEQ ID NO:60. In some embodiments, the antibody comprises oneor more of a reverted CDR-L1 comprising SEQ ID NO:62, a reverted CDR-L2comprising SEQ ID NO:30, or a reverted CDR-L3 comprising SEQ ID NO:63.In some embodiments, the antibody comprises one or more revertedframework regions, e.g., the heavy chain variable region comprisingreverted framework regions of SEQ ID NO:67.

In some embodiments, a monoclonal antibody that specifically binds to apeanut allergen comprises a heavy chain CDR1 comprising the amino acidsequence of SEQ ID NO:2 or SEQ ID NO:58, a heavy chain CDR2 comprisingthe amino acid sequence of SEQ ID NO:3 or SEQ ID NO:59, a heavy chainCDR3 comprising the amino acid sequence of SEQ ID NO:4 or SEQ ID NO:60,a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:6 orSEQ ID NO:62, a light chain CDR2 comprising the amino acid sequence ofSEQ ID NO:7 or SEQ ID NO:30, and a light chain CDR3 comprising the aminoacid sequence of SEQ ID NO:8 or SEQ ID NO:63.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:1, SEQ ID NO:57, SEQ ID NO:64, SEQ IDNO:65, SEQ ID NO:66, or SEQ ID NO:67, and comprises a light chainvariable region comprising an amino acid sequence that has at least 90%sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, or 99% sequence identity) to SEQ ID NO:5 or SEQ ID NO:61. In someembodiments, the antibody comprises a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO:57 and comprises a lightchain variable region comprising the amino acid sequence of SEQ IDNO:61. In some embodiments, the antibody comprises a heavy chainvariable region comprising the amino acid sequence of SEQ ID NO:57 andcomprises a light chain variable region comprising the amino acidsequence of SEQ ID NO:5. In some embodiments, the antibody comprises aheavy chain variable region comprising the amino acid sequence of SEQ IDNO:1 and comprises a light chain variable region comprising the aminoacid sequence of SEQ ID NO:61. In some embodiments, the antibodycomprises a heavy chain variable region comprising the amino acidsequence of SEQ ID NO:64 and comprises a light chain variable regioncomprising the amino acid sequence of SEQ ID NO:5. In some embodiments,the antibody comprises a heavy chain variable region comprising theamino acid sequence of SEQ ID NO:65 and comprises a light chain variableregion comprising the amino acid sequence of SEQ ID NO:5. In someembodiments, the antibody comprises a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO:66 and comprises a lightchain variable region comprising the amino acid sequence of SEQ ID NO:5.In some embodiments, the antibody comprises a heavy chain variableregion comprising the amino acid sequence of SEQ ID NO:67 and comprisesa light chain variable region comprising the amino acid sequence of SEQID NO:5.

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:65, and that comprises a        heavy chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:2, 59, and 4, respectively; and    -   (b) a light chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:5, and that comprises the        light chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:6, 7, and 8, respectively.

Tree Nut-Specific Antibody Sequences

In some embodiments, a monoclonal antibody that specifically binds to atree nut allergen (e.g., that specifically binds to a cashew, pistachio,almond, pine nut, pecan, walnut, hazelnut, or macadamia nut allergen)comprises heavy chain CDRs and/or light chain CDRs that are disclosed inTable 1 below. In some embodiments, a monoclonal antibody thatspecifically binds to a tree nut allergen comprises a light chainvariable region sequence and/or a heavy chain variable region sequencesequence that is disclosed in Table 1 below. In some embodiments, amonoclonal antibody that specifically binds to a tree nut allergencomprises: a heavy chain variable region comprising an amino acidsequence that has at least 75% sequence identity (e.g., at least 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequenceidentity to a heavy chain variable region sequence disclosed in Table 1and comprises a CDR1, a CDR2, and a CDR3 that is identical to the CDRsof that heavy chain variable region sequence, and a light chain variableregion comprising an amino acid sequence that has at least 70% sequenceidentity (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%,96%, 97%, 98%, or 99% sequence identity to a light chain variable regionsequence disclosed in Table 1 and comprises a CDR1, a CDR2, and a CDR3that is identical to the CDRs of that light chain variable regionsequence.

In some embodiments, a monoclonal antibody that specifically binds to atree nut allergen comprises a light chain sequence, or a portionthereof, and/or a heavy chain sequence, or a portion thereof, derivedfrom any of the following antibodies described herein: Clone PA14P3H08,Clone PA11P1D11, Clone PA11P1G10, Clone PA12P4D02, Clone PA11P1D12,Clone PA11P1F03, Clone PA11P1C04, Clone PA11P1G04, Clone PA11P1E01,Clone PA11P1C11, or Clone PA11P1C03. The amino acid sequences of theCDR, light chain variable domain (VL), and heavy chain variable domain(VH) of Clone PA14P3H08, Clone PA11P1D11, Clone PA11P1G10, ClonePA12P4D02, Clone PA11P1D12, Clone PA11P1F03, Clone PA11P1C04, ClonePA11P1G04, Clone PA11P1E01, Clone PA11P1C11, and Clone PA11P1C03 are setforth in Table 1 below.

In some embodiments, a monoclonal antibody that specifically binds to atree nut allergen comprises one or more (e.g., one, two, three, four,five, or all six) of:

-   -   (a) a heavy chain CDR1 comprising the amino acid sequence of any        one of SEQ ID NOs: 113, 167, 175, 227, 311, 318, 438, 466, 621,        665, or 692;    -   (b) a heavy chain CDR2 comprising the amino acid sequence of any        one of SEQ ID NOs: 168, 176, 200, 312, 319, 439, 539, 666, or        693;    -   (c) a heavy chain CDR3 comprising the amino acid sequence of any        one of SEQ ID NOs: 169, 177, 228, 313, 320, 440, 467, 540, 667,        or 694;    -   (d) a light chain CDR1 comprising the amino acid sequence of any        one of SEQ ID NOs: 171, 179, 230, 315, 322, 442, 469, 542, 623,        669, or 696;    -   (e) a light chain CDR2 comprising the amino acid sequence of any        one of SEQ ID NOs: 30, 94, 149, 172, 180, 196, 323, or 670; and    -   (f) a light chain CDR3 comprising the amino acid sequence of any        one of SEQ ID NOs: 173, 181, 231, 316, 324, 443, 470, 543, 624,        671, or 697.

In some embodiments, a monoclonal antibody that specifically binds to atree nut allergen comprises a heavy chain variable region comprising anamino acid sequence that comprises the sequence of, or has at least 75%sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%,95%, 96%, 97%, 98%, or 99% sequence identity) to, any one of SEQ IDNOs:166, 174, 226, 310, 317, 437, 465, 538, 620, 664, or 691. In someembodiments, a monoclonal antibody that specifically binds to a peanutallergen comprises a light chain variable region comprising an aminoacid sequence that comprises the sequence of, or has at least 75%sequence identity (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%,94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to, any one of SEQ IDNOs:170, 178, 229, 314, 321, 441, 468, 541, 622, 668, or 695. In someembodiments, a monoclonal antibody that specifically binds to a peanutallergen comprises a heavy chain variable region comprising an aminoacid sequence that comprises the sequence of, or has at least 75%sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%,95%, 96%, 97%, 98%, or 99% sequence identity to, any one of SEQ IDNOs:166, 174, 226, 310, 317, 437, 465, 538, 620, 664, or 691, andcomprises a light chain variable region comprising an amino acidsequence that comprises the sequence of, or has at least 75% sequenceidentity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%,97%, 98%, or 99% sequence identity) to, any one of SEQ ID NOs:170, 178,229, 314, 321, 441, 468, 541, 622, 668, or 695.

In some embodiments, a monoclonal antibody that specifically binds to atree nut allergen comprises:

-   -   (a) a heavy chain variable region comprising an amino acid        sequence that has at least 75% sequence identity (e.g., at least        80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%        sequence identity) to any one of SEQ ID NOs:166, 174, 226, 310,        317, 437, 465, 538, 620, 664, or 691 and that comprises a CDR1,        a CDR2, and a CDR3 that is identical to the CDRs of that SEQ ID        NO; and    -   (b) a light chain variable region comprising an amino acid        sequence that has at least 75% sequence identity (e.g., at least        80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%        sequence identity) to any one of SEQ ID NOs:170, 178, 229, 314,        321, 441, 468, 541, 622, 668, or 695 and that comprises a CDR1,        a CDR2, and a CDR3 that is identical to the CDRs of that SEQ ID        NO.

In some embodiments, the monoclonal antibody specifically binds to apecan, walnut, hazelnut, and/or macadamia nut allergen and comprises oneor more (e.g., one, two, three, four, five, or all six) of:

-   -   (a) a heavy chain CDR1 comprising the amino acid sequence of any        one of SEQ ID NOs: 113, 227, 311, 318, 665, or 692;    -   (b) a heavy chain CDR2 comprising the amino acid sequence of any        one of SEQ ID NOs: 200, 312, 319, 539, 666, or 693;    -   (c) a heavy chain CDR3 comprising the amino acid sequence of any        one of SEQ ID NOs: 228, 313, 320, 540, 667, or 694;    -   (d) a light chain CDR1 comprising the amino acid sequence of any        one of SEQ ID NOs: 230, 315, 322, 542, 669, or 696;    -   (e) a light chain CDR2 comprising the amino acid sequence of any        one of SEQ ID NOs: 94, 149, 196, 323, or 670; and    -   (f) a light chain CDR3 comprising the amino acid sequence of any        one of SEQ ID NOs: 231, 316, 324, 543, 671, or 697.

In some embodiments, a monoclonal antibody that specifically binds to apecan, walnut, hazelnut, and/or macadamia nut allergen comprises a heavychain variable region comprising an amino acid sequence that comprisesthe sequence of, or has at least 75% sequence identity (e.g., at least80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequenceidentity) to, any one of SEQ ID NOs: 226, 310, 317, 538, 664, or 691. Insome embodiments, a monoclonal antibody that specifically binds to apecan, walnut, hazelnut, and/or macadamia nut allergen comprises a lightchain variable region comprising an amino acid sequence that comprisesthe sequence of, or has at least 75% sequence identity (e.g., at least75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to, any one of SEQ ID NOs: 229, 314, 321, 541, 668,or 695. In some embodiments, a monoclonal antibody that specificallybinds to a pecan, walnut, hazelnut, and/or macadamia nut allergencomprises a heavy chain variable region comprising an amino acidsequence that comprises the sequence of, or has at least 75% sequenceidentity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%,97%, 98%, or 99% sequence identity to, any one of SEQ ID NOs: 226, 310,317, 437, 538, 664, or 691, and comprises a light chain variable regioncomprising an amino acid sequence that comprises the sequence of, or hasat least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%,93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to, any one ofSEQ ID NOs: 229, 314, 321, 541, 668, or 695.

In some embodiments, the monoclonal antibody specifically binds to acashew and/or pistachio allergen and comprises one or more (e.g., one,two, three, four, five, or all six) of:

-   -   (a) a heavy chain CDR1 comprising the amino acid sequence of any        one of SEQ ID NOs: 167, 175, 227, 438, 466, or 621;    -   (b) a heavy chain CDR2 comprising the amino acid sequence of any        one of SEQ ID NOs: 168, 176, 200, or 439;    -   (c) a heavy chain CDR3 comprising the amino acid sequence of any        one of SEQ ID NOs: 169, 177, 440, or 467;    -   (d) a light chain CDR1 comprising the amino acid sequence of any        one of SEQ ID NOs: 171, 179, 442, 469, or 623;    -   (e) a light chain CDR2 comprising the amino acid sequence of any        one of SEQ ID NOs: 30, 149, 172, or 180; and    -   (f) a light chain CDR3 comprising the amino acid sequence of any        one of SEQ ID NOs: 173, 181, 443, 470, or 624.

In some embodiments, a monoclonal antibody that specifically binds to acashew and/or pistachio allergen comprises a heavy chain variable regioncomprising an amino acid sequence that comprises the sequence of, or hasat least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%,93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to, any one ofSEQ ID NOs:166, 174, 437, 465, or 620. In some embodiments, a monoclonalantibody that specifically binds to a cashew and/or pistachio allergencomprises a light chain variable region comprising an amino acidsequence that comprises the sequence of, or has at least 75% sequenceidentity (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%,96%, 97%, 98%, or 99% sequence identity) to, any one of SEQ ID NOs:170,178, 441, 468, or 622. In some embodiments, a monoclonal antibody thatspecifically binds to a cashew and/or pistachio allergen comprises aheavy chain variable region comprising an amino acid sequence thatcomprises the sequence of, or has at least 75% sequence identity (e.g.,at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity to, any one of SEQ ID NOs:166, 174, 437, 465, or 620,and comprises a light chain variable region comprising an amino acidsequence that comprises the sequence of, or has at least 75% sequenceidentity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%,97%, 98%, or 99% sequence identity) to, any one of SEQ ID NOs:170, 178,441, 468, or 622.

In some embodiments, a monoclonal antibody that specifically binds to atree nut allergen comprises a heavy chain CDR1-3 and a light chainCDR1-3 comprising the amino acid sequences of:

-   -   (a) SEQ ID NOs:692, 693, 694, 696, 94, and 697, respectively; or    -   (b) SEQ ID NOs:318, 319, 320, 322, 323, and 324, respectively;        or    -   (c) SEQ ID NOs:227, 200, 228, 230, 149, and 231, respectively;        or    -   (d) SEQ ID NOs:113, 539, 540, 542, 196, and 543, respectively;        or    -   (e) SEQ ID NOs:311, 312, 313, 315, 94, and 316, respectively; or    -   (f) SEQ ID NOs:665, 666, 667, 669, 670, and 671, respectively;        or    -   (g) SEQ ID NOs:466, 200, 467, 469, 149, and 470, respectively;        or    -   (h) SEQ ID NOs:167, 168, 169, 171, 172, and 173, respectively;        or    -   (i) SEQ ID NOs:621, 176, 177, 623, 180, and 624, respectively;        or    -   (j) SEQ ID NOs:175, 176, 177, 179, 180, and 181, respectively;        or    -   (k) SEQ ID NOs:438, 439, 440, 442, 30, and 443, respectively.

Clone PA14P3H08

In some embodiments, a monoclonal antibody that specifically binds to atree nut allergen comprises a heavy chain CDR1 comprising the amino acidsequence of SEQ ID NO:692, a heavy chain CDR2 comprising the amino acidsequence of SEQ ID NO:693, a heavy chain CDR3 comprising the amino acidsequence of SEQ ID NO:694, a light chain CDR1 comprising the amino acidsequence of SEQ ID NO:696, a light chain CDR2 comprising the amino acidsequence of SEQ ID NO:94, and a light chain CDR3 comprising the aminoacid sequence of SEQ ID NO:697. In some embodiments, the monoclonalantibody binds to two or more tree nut allergens. In some embodiments,the monoclonal antibody binds to a pecan, walnut, hazelnut, and/ormacadamia nut allergen.

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:691, and that comprises a        heavy chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:692, 693, and 694, respectively; and    -   (b) a light chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:695, and that comprises        the light chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:696, 94, and 697, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:691, and comprises a light chainvariable region comprising an amino acid sequence that has at least 90%sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, or 99% sequence identity) to SEQ ID NO:695. In some embodiments,the antibody comprises a heavy chain variable region comprising theamino acid sequence of SEQ ID NO:691 and comprises a light chainvariable region comprising the amino acid sequence of SEQ ID NO:695.

In some embodiments, the antibody is an antibody that competes forbinding with an antibody as described herein (e.g., an antibodycomprising a heavy chain CDR1-3 and a light chain CDR1-3 comprising theamino acid sequences of SEQ ID NOs:692, 693, 694, 696, 95, and 697,respectively, or an antibody comprising a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO:691 and furthercomprising a light chain variable region comprising the amino acidsequence of SEQ ID NO:695).

Clone PA11P1D11

In some embodiments, a monoclonal antibody that specifically binds to atree nut allergen comprises a heavy chain CDR1 comprising the amino acidsequence of SEQ ID NO:318, a heavy chain CDR2 comprising the amino acidsequence of SEQ ID NO:319, a heavy chain CDR3 comprising the amino acidsequence of SEQ ID NO:320, a light chain CDR1 comprising the amino acidsequence of SEQ ID NO:322, a light chain CDR2 comprising the amino acidsequence of SEQ ID NO:323, and a light chain CDR3 comprising the aminoacid sequence of SEQ ID NO:324. In some embodiments, the monoclonalantibody binds to two or more tree nut allergens. In some embodiments,the monoclonal antibody binds to a pecan, walnut, hazelnut, and/ormacadamia nut allergen.

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:317, and that comprises a        heavy chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:318, 319, and 320, respectively; and    -   (b) a light chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:321, and that comprises        the light chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:322, 323, and 324, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:317, and comprises a light chainvariable region comprising an amino acid sequence that has at least 90%sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, or 99% sequence identity) to SEQ ID NO:321. In some embodiments,the antibody comprises a heavy chain variable region comprising theamino acid sequence of SEQ ID NO:317 and comprises a light chainvariable region comprising the amino acid sequence of SEQ ID NO:321.

In some embodiments, the antibody is an antibody that competes forbinding with an antibody as described herein (e.g., an antibodycomprising a heavy chain CDR1-3 and a light chain CDR1-3 comprising theamino acid sequences of SEQ ID NOs:318, 319, 320, 322, 323, and 324,respectively, or an antibody comprising a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO:317 and furthercomprising a light chain variable region comprising the amino acidsequence of SEQ ID NO:321).

Clone PA11P1G10

In some embodiments, a monoclonal antibody that specifically binds to atree nut allergen comprises a heavy chain CDR1 comprising the amino acidsequence of SEQ ID NO:227, a heavy chain CDR2 comprising the amino acidsequence of SEQ ID NO:200, a heavy chain CDR3 comprising the amino acidsequence of SEQ ID NO:228, a light chain CDR1 comprising the amino acidsequence of SEQ ID NO:230, a light chain CDR2 comprising the amino acidsequence of SEQ ID NO:149, and a light chain CDR3 comprising the aminoacid sequence of SEQ ID NO:231. In some embodiments, the monoclonalantibody binds to two or more tree nut allergens. In some embodiments,the monoclonal antibody binds to a pecan, walnut, hazelnut, and/ormacadamia nut allergen.

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:226, and that comprises a        heavy chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:227, 200, and 228, respectively; and    -   (b) a light chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:229, and that comprises        the light chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:230, 149, and 231, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:226, and comprises a light chainvariable region comprising an amino acid sequence that has at least 90%sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, or 99% sequence identity) to SEQ ID NO:229. In some embodiments,the antibody comprises a heavy chain variable region comprising theamino acid sequence of SEQ ID NO:226 and comprises a light chainvariable region comprising the amino acid sequence of SEQ ID NO:229.

In some embodiments, the antibody is an antibody that competes forbinding with an antibody as described herein (e.g., an antibodycomprising a heavy chain CDR1-3 and a light chain CDR1-3 comprising theamino acid sequences of SEQ ID NOs:227, 200, 228, 230, 149, and 231,respectively, or an antibody comprising a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO:226 and furthercomprising a light chain variable region comprising the amino acidsequence of SEQ ID NO:229).

Clone PA12P4D02

In some embodiments, a monoclonal antibody that specifically binds to atree nut allergen comprises a heavy chain CDR1 comprising the amino acidsequence of SEQ ID NO:113, a heavy chain CDR2 comprising the amino acidsequence of SEQ ID NO:539, a heavy chain CDR3 comprising the amino acidsequence of SEQ ID NO:540, a light chain CDR1 comprising the amino acidsequence of SEQ ID NO:542, a light chain CDR2 comprising the amino acidsequence of SEQ ID NO:196, and a light chain CDR3 comprising the aminoacid sequence of SEQ ID NO:543. In some embodiments, the monoclonalantibody binds to two or more tree nut allergens. In some embodiments,the monoclonal antibody binds to a pecan, walnut, hazelnut, and/ormacadamia nut allergen.

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:538, and that comprises a        heavy chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:113, 539, and 540, respectively; and    -   (b) a light chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:541, and that comprises        the light chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:542, 196, and 543, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:538, and comprises a light chainvariable region comprising an amino acid sequence that has at least 90%sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, or 99% sequence identity) to SEQ ID NO:541. In some embodiments,the antibody comprises a heavy chain variable region comprising theamino acid sequence of SEQ ID NO:538 and comprises a light chainvariable region comprising the amino acid sequence of SEQ ID NO:541.

In some embodiments, the antibody is an antibody that competes forbinding with an antibody as described herein (e.g., an antibodycomprising a heavy chain CDR1-3 and a light chain CDR1-3 comprising theamino acid sequences of SEQ ID NOs:113, 539, 540, 542, 196, and 543,respectively, or an antibody comprising a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO:538 and furthercomprising a light chain variable region comprising the amino acidsequence of SEQ ID NO:541).

Clone PA11P1D12

In some embodiments, a monoclonal antibody that specifically binds to atree nut allergen comprises a heavy chain CDR1 comprising the amino acidsequence of SEQ ID NO:692, a heavy chain CDR2 comprising the amino acidsequence of SEQ ID NO:693, a heavy chain CDR3 comprising the amino acidsequence of SEQ ID NO:694, a light chain CDR1 comprising the amino acidsequence of SEQ ID NO:696, a light chain CDR2 comprising the amino acidsequence of SEQ ID NO:94, and a light chain CDR3 comprising the aminoacid sequence of SEQ ID NO:697. In some embodiments, the monoclonalantibody binds to two or more tree nut allergens. In some embodiments,the monoclonal antibody binds to a pecan, walnut, hazelnut, and/ormacadamia nut allergen.

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:310, and that comprises a        heavy chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:311, 312, and 313, respectively; and    -   (b) a light chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:314, and that comprises        the light chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:315, 94, and 316, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:310, and comprises a light chainvariable region comprising an amino acid sequence that has at least 90%sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, or 99% sequence identity) to SEQ ID NO:314. In some embodiments,the antibody comprises a heavy chain variable region comprising theamino acid sequence of SEQ ID NO:310 and comprises a light chainvariable region comprising the amino acid sequence of SEQ ID NO:314.

In some embodiments, the antibody is an antibody that competes forbinding with an antibody as described herein (e.g., an antibodycomprising a heavy chain CDR1-3 and a light chain CDR1-3 comprising theamino acid sequences of SEQ ID NOs:311, 312, 313, 315, 94, and 316,respectively, or an antibody comprising a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO:310 and furthercomprising a light chain variable region comprising the amino acidsequence of SEQ ID NO:314).

Clone PA11P1F03

In some embodiments, a monoclonal antibody that specifically binds to atree nut allergen comprises a heavy chain CDR1 comprising the amino acidsequence of SEQ ID NO:665, a heavy chain CDR2 comprising the amino acidsequence of SEQ ID NO:666, a heavy chain CDR3 comprising the amino acidsequence of SEQ ID NO:667, a light chain CDR1 comprising the amino acidsequence of SEQ ID NO:669, a light chain CDR2 comprising the amino acidsequence of SEQ ID NO:670, and a light chain CDR3 comprising the aminoacid sequence of SEQ ID NO:671. In some embodiments, the monoclonalantibody binds to two or more tree nut allergens. In some embodiments,the monoclonal antibody binds to a pecan, walnut, hazelnut, and/ormacadamia nut allergen.

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:664, and that comprises a        heavy chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:665, 666, and 667, respectively; and    -   (b) a light chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:668, and that comprises        the light chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:669, 670, and 671, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:664, and comprises a light chainvariable region comprising an amino acid sequence that has at least 90%sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, or 99% sequence identity) to SEQ ID NO:668. In some embodiments,the antibody comprises a heavy chain variable region comprising theamino acid sequence of SEQ ID NO:664 and comprises a light chainvariable region comprising the amino acid sequence of SEQ ID NO:668.

In some embodiments, the antibody is an antibody that competes forbinding with an antibody as described herein (e.g., an antibodycomprising a heavy chain CDR1-3 and a light chain CDR1-3 comprising theamino acid sequences of SEQ ID NOs:665, 666, 667, 669, 670, and 671,respectively, or an antibody comprising a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO:664 and furthercomprising a light chain variable region comprising the amino acidsequence of SEQ ID NO:668).

Clone PA11P1C04

In some embodiments, a monoclonal antibody that specifically binds to atree nut allergen comprises a heavy chain CDR1 comprising the amino acidsequence of SEQ ID NO:466, a heavy chain CDR2 comprising the amino acidsequence of SEQ ID NO:200, a heavy chain CDR3 comprising the amino acidsequence of SEQ ID NO:467, a light chain CDR1 comprising the amino acidsequence of SEQ ID NO:469, a light chain CDR2 comprising the amino acidsequence of SEQ ID NO:149, and a light chain CDR3 comprising the aminoacid sequence of SEQ ID NO:470. In some embodiments, the monoclonalantibody binds to two or more tree nut allergens. In some embodiments,the monoclonal antibody binds to a cashew and/or a pistachio allergen.

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:465, and that comprises a        heavy chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:466, 200, and 467, respectively; and    -   (b) a light chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:468, and that comprises        the light chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:469, 149, and 470, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:465, and comprises a light chainvariable region comprising an amino acid sequence that has at least 90%sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, or 99% sequence identity) to SEQ ID NO:468. In some embodiments,the antibody comprises a heavy chain variable region comprising theamino acid sequence of SEQ ID NO:465 and comprises a light chainvariable region comprising the amino acid sequence of SEQ ID NO:468.

In some embodiments, the antibody is an antibody that competes forbinding with an antibody as described herein (e.g., an antibodycomprising a heavy chain CDR1-3 and a light chain CDR1-3 comprising theamino acid sequences of SEQ ID NOs:466, 200, 467, 469, 149, and 470,respectively, or an antibody comprising a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO:465 and furthercomprising a light chain variable region comprising the amino acidsequence of SEQ ID NO:468).

Clone PA11P1G04

In some embodiments, a monoclonal antibody that specifically binds to atree nut allergen comprises a heavy chain CDR1 comprising the amino acidsequence of SEQ ID NO:167, a heavy chain CDR2 comprising the amino acidsequence of SEQ ID NO:168, a heavy chain CDR3 comprising the amino acidsequence of SEQ ID NO:169, a light chain CDR1 comprising the amino acidsequence of SEQ ID NO:171, a light chain CDR2 comprising the amino acidsequence of SEQ ID NO:172, and a light chain CDR3 comprising the aminoacid sequence of SEQ ID NO:173. In some embodiments, the monoclonalantibody binds to two or more tree nut allergens. In some embodiments,the monoclonal antibody binds to a cashew and/or a pistachio allergen.

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:166, and that comprises a        heavy chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:167, 168, and 169, respectively; and    -   (b) a light chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:170, and that comprises        the light chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:171, 172, and 173, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:166, and comprises a light chainvariable region comprising an amino acid sequence that has at least 90%sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, or 99% sequence identity) to SEQ ID NO:170. In some embodiments,the antibody comprises a heavy chain variable region comprising theamino acid sequence of SEQ ID NO:166 and comprises a light chainvariable region comprising the amino acid sequence of SEQ ID NO:170.

In some embodiments, the antibody is an antibody that competes forbinding with an antibody as described herein (e.g., an antibodycomprising a heavy chain CDR1-3 and a light chain CDR1-3 comprising theamino acid sequences of SEQ ID NOs:167, 168, 169, 171, 172, and 173,respectively, or an antibody comprising a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO:166 and furthercomprising a light chain variable region comprising the amino acidsequence of SEQ ID NO:170).

Clone PA11P1E01

In some embodiments, a monoclonal antibody that specifically binds to atree nut allergen comprises a heavy chain CDR1 comprising the amino acidsequence of SEQ ID NO:621, a heavy chain CDR2 comprising the amino acidsequence of SEQ ID NO:176, a heavy chain CDR3 comprising the amino acidsequence of SEQ ID NO:177, a light chain CDR1 comprising the amino acidsequence of SEQ ID NO:623, a light chain CDR2 comprising the amino acidsequence of SEQ ID NO:180, and a light chain CDR3 comprising the aminoacid sequence of SEQ ID NO:624. In some embodiments, the monoclonalantibody binds to two or more tree nut allergens. In some embodiments,the monoclonal antibody binds to a cashew and/or a pistachio allergen.

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:620, and that comprises a        heavy chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:621, 176, and 177, respectively; and    -   (b) a light chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:622, and that comprises        the light chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:623, 180, and 624, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:620, and comprises a light chainvariable region comprising an amino acid sequence that has at least 90%sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, or 99% sequence identity) to SEQ ID NO:622. In some embodiments,the antibody comprises a heavy chain variable region comprising theamino acid sequence of SEQ ID NO:620 and comprises a light chainvariable region comprising the amino acid sequence of SEQ ID NO:622.

In some embodiments, the antibody is an antibody that competes forbinding with an antibody as described herein (e.g., an antibodycomprising a heavy chain CDR1-3 and a light chain CDR1-3 comprising theamino acid sequences of SEQ ID NOs:621, 176, 177, 623, 180, and 624,respectively, or an antibody comprising a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO:620 and furthercomprising a light chain variable region comprising the amino acidsequence of SEQ ID NO:622).

Clone PA11P1C11

In some embodiments, a monoclonal antibody that specifically binds to atree nut allergen comprises a heavy chain CDR1 comprising the amino acidsequence of SEQ ID NO:175, a heavy chain CDR2 comprising the amino acidsequence of SEQ ID NO:176, a heavy chain CDR3 comprising the amino acidsequence of SEQ ID NO:177, a light chain CDR1 comprising the amino acidsequence of SEQ ID NO:179, a light chain CDR2 comprising the amino acidsequence of SEQ ID NO:180, and a light chain CDR3 comprising the aminoacid sequence of SEQ ID NO:181. In some embodiments, the monoclonalantibody binds to two or more tree nut allergens. In some embodiments,the monoclonal antibody binds to a cashew and/or a pistachio allergen.

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:174, and that comprises a        heavy chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:175, 176, and 177, respectively; and    -   (b) a light chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:178, and that comprises        the light chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:179, 180, and 181, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:174, and comprises a light chainvariable region comprising an amino acid sequence that has at least 90%sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, or 99% sequence identity) to SEQ ID NO:178. In some embodiments,the antibody comprises a heavy chain variable region comprising theamino acid sequence of SEQ ID NO:174 and comprises a light chainvariable region comprising the amino acid sequence of SEQ ID NO:178.

In some embodiments, the antibody is an antibody that competes forbinding with an antibody as described herein (e.g., an antibodycomprising a heavy chain CDR1-3 and a light chain CDR1-3 comprising theamino acid sequences of SEQ ID NOs:175, 176, 177, 179, 180, and 181,respectively, or an antibody comprising a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO:174 and furthercomprising a light chain variable region comprising the amino acidsequence of SEQ ID NO:178).

Clone PA11P1C03

In some embodiments, a monoclonal antibody that specifically binds to atree nut allergen comprises a heavy chain CDR1 comprising the amino acidsequence of SEQ ID NO:438, a heavy chain CDR2 comprising the amino acidsequence of SEQ ID NO:439, a heavy chain CDR3 comprising the amino acidsequence of SEQ ID NO:440, a light chain CDR1 comprising the amino acidsequence of SEQ ID NO:442, a light chain CDR2 comprising the amino acidsequence of SEQ ID NO:30, and a light chain CDR3 comprising the aminoacid sequence of SEQ ID NO:443. In some embodiments, the monoclonalantibody binds to two or more tree nut allergens. In some embodiments,the monoclonal antibody binds to a cashew and/or a pistachio allergen.

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:437, and that comprises a        heavy chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:438, 439, and 440, respectively; and    -   (b) a light chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:441, and that comprises        the light chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:442, 30, and 443, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:437, and comprises a light chainvariable region comprising an amino acid sequence that has at least 90%sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, or 99% sequence identity) to SEQ ID NO:441. In some embodiments,the antibody comprises a heavy chain variable region comprising theamino acid sequence of SEQ ID NO:437 and comprises a light chainvariable region comprising the amino acid sequence of SEQ ID NO:441.

In some embodiments, the antibody is an antibody that competes forbinding with an antibody as described herein (e.g., an antibodycomprising a heavy chain CDR1-3 and a light chain CDR1-3 comprising theamino acid sequences of SEQ ID NOs:438, 439, 440, 442, 30, and 443,respectively, or an antibody comprising a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO:437 and furthercomprising a light chain variable region comprising the amino acidsequence of SEQ ID NO:441).

Antibodies that Specifically Bind to Milk Allergens

In some embodiments, the monoclonal antibody or antigen-binding portionthereof specifically binds to a milk allergen (e.g., cow's milkallergen). In some embodiments, a monoclonal antibody that specificallybinds to a milk allergen comprises heavy chain CDRs and/or light chainCDRs that are disclosed in Table 1 below. In some embodiments, amonoclonal antibody that specifically binds to a milk allergen comprisesa light chain variable region sequence and/or a heavy chain variableregion sequence sequence that is disclosed in Table 1 below. In someembodiments, a monoclonal antibody that specifically binds to a milkallergen comprises: a heavy chain variable region comprising an aminoacid sequence that has at least 75% sequence identity (e.g., at least80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequenceidentity to a heavy chain variable region sequence disclosed in Table 1and comprises a CDR1, a CDR2, and a CDR3 that is identical to the CDRsof that heavy chain variable region sequence, and a light chain variableregion comprising an amino acid sequence that has at least 70% sequenceidentity (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%,96%, 97%, 98%, or 99% sequence identity to a light chain variable regionsequence disclosed in Table 1 and comprises a CDR1, a CDR2, and a CDR3that is identical to the CDRs of that light chain variable regionsequence.

In some embodiments, a monoclonal antibody that specifically binds to amilk allergen comprises a light chain sequence, or a portion thereof,and/or a heavy chain sequence, or a portion thereof, derived from any ofthe following antibodies described herein: Clone PA01P2C05, ClonePA01P2B03, Clone PA01P2A12, Clone PA01P2C12, Clone PA01P2E10, ClonePA01P2C09, Clone PA01P2D06, Clone PA01P2E08, Clone PA01P2A05, ClonePA01P2B04, Clone PA01P2E05, Clone PA01P2D04, Clone PA01P2B12, ClonePA01P2D11, Clone PA01P2B10, Clone PA01P2D10, Clone PA01P2D09, ClonePA01P2B05, Clone PA01P4C11, Clone PA01P3E08, Clone PA01P2E06, ClonePA01P2E07, Clone PA01P2G07, Clone PA01P2B09, Clone PA01P2C04, or ClonePA01P2H08. The amino acid sequences of the CDR, light chain variabledomain (VL), and heavy chain variable domain (VH) of Clone PA01P2C05,Clone PA01P2B03, Clone PA01P2A12, Clone PA01P2C12, Clone PA01P2E10,Clone PA01P2C09, Clone PA01P2D06, Clone PA01P2E08, Clone PA01P2A05,Clone PA01P2B04, Clone PA01P2E05, Clone PA01P2D04, Clone PA01P2B12,Clone PA01P2D11, Clone PA01P2B10, Clone PA01P2D10, Clone PA01P2D09,Clone PA01P2B05, Clone PA01P4C11, Clone PA01P3E08, Clone PA01P2E06,Clone PA01P2E07, Clone PA01P2G07, Clone PA01P2B09, Clone PA01P2C04, andClone PA01P2H08 are set forth in Table 1 below.

In some embodiments, a monoclonal antibody that specifically binds to apeanut allergen comprises one or more (e.g., one, two, three, four,five, or all six) of:

-   -   (a) a heavy chain CDR1 comprising the amino acid sequence of any        one of SEQ ID NOs: 121, 750, 757, 765, 772, 779, 785, 793, 800,        807, 814, 821, 833, 838, 846, 853, 860, 868, 874, 881, 889, 895,        903, 911, 918, or 926;    -   (b) a heavy chain CDR2 comprising the amino acid sequence of any        one of SEQ ID NOs: 378, 532, 751, 758, 766, 773, 786, 794, 801,        808, 815, 822, 826, 839, 847, 854, 861, 875, 882, 890, 896, 904,        912, 919, or 927;    -   (c) a heavy chain CDR3 comprising the amino acid sequence of any        one of SEQ ID NOs: 752, 759, 767, 774, 780, 787, 795, 802, 809,        816, 827, 840, 848, 855, 862, 869, 876, 883, 891, 897, 905, 913,        920, or 928;    -   (d) a light chain CDR1 comprising the amino acid sequence of any        one of SEQ ID NOs: 523, 754, 761, 769, 776, 782, 789, 797, 804,        811, 818, 829, 835, 842, 850, 857, 864, 871, 878, 885, 899, 907,        915, 922, or 930;    -   (e) a light chain CDR2 comprising the amino acid sequence of any        one of SEQ ID NOs: 22, 30, 94, 110, 149, 186, 196, 389, 404,        509, 662, 682, 762, 790, 830, 843, 865, 886, 900, 908, 923, or        931; and    -   (f) a light chain CDR3 comprising the amino acid sequence of any        one of SEQ ID NOs: 755, 763, 770, 777, 783, 791, 798, 805, 812,        819, 824, 831, 836, 844, 851, 858, 866, 872, 879, 887, 893, 901,        909, 916, 924, or 932.

In some embodiments, a monoclonal antibody that specifically binds to amilk allergen comprises a heavy chain CDR1-3 and a light chain CDR1-3comprising the amino acid sequences of:

-   -   (a) SEQ ID NOs:750, 751, 752, 754, 196, and 755, respectively;        or    -   (b) SEQ ID NOs:757, 758, 759, 761, 762, and 763, respectively;        or    -   (c) SEQ ID NOs:765, 766, 767, 769, 30, and 770, respectively; or    -   (d) SEQ ID NOs:772, 773, 774, 776, 389, and 777, respectively;        or    -   (e) SEQ ID NOs:779, 532, 780, 782, 404, and 783, respectively;        or    -   (f) SEQ ID NOs:785, 786, 787, 789, 790, and 791, respectively;        or    -   (g) SEQ ID NOs:793, 794, 795, 797, 30, and 798, respectively; or    -   (h) SEQ ID NOs:800, 801, 802, 804, 110, and 805, respectively;        or    -   (i) SEQ ID NOs:807, 808, 809, 811, 509, and 812, respectively;        or    -   (j) SEQ ID NOs:814, 815, 816, 818, 94, and 819, respectively; or    -   (k) SEQ ID NOs:821, 822, 787, 776, 790, and 824, respectively;        or    -   (l) SEQ ID NOs:121, 826, 827, 829, 830, and 831, respectively;        or    -   (m) SEQ ID NOs:833, 826, 827, 835, 149, and 836, respectively;        or    -   (n) SEQ ID NOs:838, 839, 840, 842, 843, and 844, respectively;        or    -   (o) SEQ ID NOs:846, 847, 848, 850, 196, and 851, respectively;        or    -   (p) SEQ ID NOs:853, 854, 855, 857, 662, and 858, respectively;        or    -   (q) SEQ ID NOs:860, 861, 862, 864, 865, and 866, respectively;        or    -   (r) SEQ ID NOs:868, 378, 869, 871, 682, and 872, respectively;        or    -   (s) SEQ ID NOs:874, 875, 876, 878, 22, and 879, respectively; or    -   (t) SEQ ID NOs:881, 882, 883, 885, 886, and 887, respectively;        or    -   (u) SEQ ID NOs:889, 890, 891, 523, 762, and 893, respectively;        or    -   (v) SEQ ID NOs:895, 896, 897, 899, 900, and 901, respectively;        or    -   (w) SEQ ID NOs:903, 904, 905, 907, 908, and 909, respectively;        or    -   (x) SEQ ID NOs:911, 912, 913, 915, 149, and 916, respectively;        or    -   (y) SEQ ID NOs:918, 919, 920, 922, 923, and 924, respectively;        or    -   (z) SEQ ID NOs:926, 927, 928, 930, 931, and 932, respectively.

In some embodiments, a monoclonal antibody that specifically binds to amilk allergen comprises a heavy chain variable region comprising anamino acid sequence that has at least 75% sequence identity (e.g., atleast 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to any one of SEQ ID NOs:749, 756, 764, 771, 778,784, 792, 799, 806, 813, 820, 825, 832, 837, 845, 852, 859, 867, 873,880, 888, 894, 902, 910, 917, or 925. In some embodiments, a monoclonalantibody that specifically binds to a milk allergen comprises a lightchain variable region comprising an amino acid sequence that has atleast 75% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 91%,92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity) to any oneof SEQ ID NOs:753, 760, 768, 775, 781, 788, 796, 803, 810, 817, 823,828, 834, 841, 849, 856, 863, 870, 877, 884, 892, 898, 906, 914, 921, or929. In some embodiments, a monoclonal antibody that specifically bindsto a milk allergen comprises a heavy chain variable region comprising anamino acid sequence that has at least 75% sequence identity (e.g., atleast 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to any one of SEQ ID NOs:749, 756, 764, 771, 778,784, 792, 799, 806, 813, 820, 825, 832, 837, 845, 852, 859, 867, 873,880, 888, 894, 902, 910, 917, or 925, and comprises a light chainvariable region comprising an amino acid sequence that has at least 75%sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%, 93%, 94%,95%, 96%, 97%, 98%, or 99% sequence identity) to any one of SEQ IDNOs:753, 760, 768, 775, 781, 788, 796, 803, 810, 817, 823, 828, 834,841, 849, 856, 863, 870, 877, 884, 892, 898, 906, 914, 921, or 929.

In some embodiments, a monoclonal antibody that specifically binds to amilk allergen comprises a heavy chain variable region comprising anamino acid sequence that has at least 75% sequence identity (e.g., atleast 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to any one of SEQ ID NOs:749, 756, 764, 771, 778,784, 792, 799, 806, 813, 820, 825, 832, 837, 845, 852, 859, 867, 873,880, 888, 894, 902, 910, 917, or 925 and comprises a CDR1, a CDR2, and aCDR3 that is identical to the CDRs of that SEQ ID NO. In someembodiments, a monoclonal antibody that specifically binds to a milkallergen comprises a light chain variable region comprising an aminoacid sequence that has at least 70% sequence identity (e.g., at least75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to any one of SEQ ID NOs:753, 760, 768, 775, 781,788, 796, 803, 810, 817, 823, 828, 834, 841, 849, 856, 863, 870, 877,884, 892, 898, 906, 914, 921, or 929 and comprises a CDR1, a CDR2, and aCDR3 that is identical to the CDRs of that SEQ ID NO.

In some embodiments, a monoclonal antibody that specifically binds to amilk allergen comprises:

-   -   (a) a heavy chain variable region comprising an amino acid        sequence that has at least 90% (e.g., at least 75%, 80%, 85%,        90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence        identity to SEQ ID NO:749 or that has the sequence of SEQ ID        NO:749, and a light chain variable region comprising an amino        acid sequence that has at least 90% (e.g., at least 75%, 80%,        85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%)        sequence identity to SEQ ID NO:753 or that has the sequence of        SEQ ID NO:753; or    -   (b) a heavy chain variable region comprising an amino acid        sequence that has at least 90% (e.g., at least 75%, 80%, 85%,        90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence        identity to SEQ ID NO:756 or that has the sequence of SEQ ID        NO:756, and a light chain variable region comprising an amino        acid sequence that has at least 90% (e.g., at least 75%, 80%,        85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%)        sequence identity to SEQ ID NO:760 or that has the sequence of        SEQ ID NO:760; or    -   (c) a heavy chain variable region comprising an amino acid        sequence that has at least 90% (e.g., at least 75%, 80%, 85%,        90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence        identity to SEQ ID NO:764 or that has the sequence of SEQ ID        NO:764, and a light chain variable region comprising an amino        acid sequence that has at least 90% (e.g., at least 75%, 80%,        85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%)        sequence identity to SEQ ID NO:768 or that has the sequence of        SEQ ID NO:768; or    -   (d) a heavy chain variable region comprising an amino acid        sequence that has at least 90% (e.g., at least 75%, 80%, 85%,        90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence        identity to SEQ ID NO:771 or that has the sequence of SEQ ID        NO:771, and a light chain variable region comprising an amino        acid sequence that has at least 90% (e.g., at least 75%, 80%,        85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%)        sequence identity to SEQ ID NO:775 or that has the sequence of        SEQ ID NO:775; or    -   (e) a heavy chain variable region comprising an amino acid        sequence that has at least 90% (e.g., at least 75%, 80%, 85%,        90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence        identity to SEQ ID NO:778 or that has the sequence of SEQ ID        NO:778, and a light chain variable region comprising an amino        acid sequence that has at least 90% (e.g., at least 75%, 80%,        85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%)        sequence identity to SEQ ID NO:781 or that has the sequence of        SEQ ID NO:781; or    -   (f) a heavy chain variable region comprising an amino acid        sequence that has at least 90% (e.g., at least 75%, 80%, 85%,        90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence        identity to SEQ ID NO:784 or that has the sequence of SEQ ID        NO:784, and a light chain variable region comprising an amino        acid sequence that has at least 90% (e.g., at least 75%, 80%,        85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%)        sequence identity to SEQ ID NO:788 or that has the sequence of        SEQ ID NO:788; or    -   (g) a heavy chain variable region comprising an amino acid        sequence that has at least 90% (e.g., at least 75%, 80%, 85%,        90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence        identity to SEQ ID NO:792 or that has the sequence of SEQ ID        NO:792, and a light chain variable region comprising an amino        acid sequence that has at least 90% (e.g., at least 75%, 80%,        85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%)        sequence identity to SEQ ID NO:796 or that has the sequence of        SEQ ID NO:796; or    -   (h) a heavy chain variable region comprising an amino acid        sequence that has at least 90% (e.g., at least 75%, 80%, 85%,        90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence        identity to SEQ ID NO:799 or that has the sequence of SEQ ID        NO:799, and a light chain variable region comprising an amino        acid sequence that has at least 90% (e.g., at least 75%, 80%,        85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%)        sequence identity to SEQ ID NO:803 or that has the sequence of        SEQ ID NO:803; or    -   (i) a heavy chain variable region comprising an amino acid        sequence that has at least 90% (e.g., at least 75%, 80%, 85%,        90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence        identity to SEQ ID NO:806 or that has the sequence of SEQ ID        NO:806, and a light chain variable region comprising an amino        acid sequence that has at least 90% (e.g., at least 75%, 80%,        85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%)        sequence identity to SEQ ID NO:810 or that has the sequence of        SEQ ID NO:810; or    -   (j) a heavy chain variable region comprising an amino acid        sequence that has at least 90% (e.g., at least 75%, 80%, 85%,        90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence        identity to SEQ ID NO:813 or that has the sequence of SEQ ID        NO:813, and a light chain variable region comprising an amino        acid sequence that has at least 90% (e.g., at least 75%, 80%,        85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%)        sequence identity to SEQ ID NO:817 or that has the sequence of        SEQ ID NO:817; or    -   (k) a heavy chain variable region comprising an amino acid        sequence that has at least 90% (e.g., at least 75%, 80%, 85%,        90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence        identity to SEQ ID NO:820 or that has the sequence of SEQ ID        NO:820, and a light chain variable region comprising an amino        acid sequence that has at least 90% (e.g., at least 75%, 80%,        85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%)        sequence identity to SEQ ID NO:823 or that has the sequence of        SEQ ID NO:823; or    -   (l) a heavy chain variable region comprising an amino acid        sequence that has at least 90% (e.g., at least 75%, 80%, 85%,        90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence        identity to SEQ ID NO:825 or that has the sequence of SEQ ID        NO:825, and a light chain variable region comprising an amino        acid sequence that has at least 90% (e.g., at least 75%, 80%,        85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%)        sequence identity to SEQ ID NO:828 or that has the sequence of        SEQ ID NO:828; or    -   (m) a heavy chain variable region comprising an amino acid        sequence that has at least 90% (e.g., at least 75%, 80%, 85%,        90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence        identity to SEQ ID NO:832 or that has the sequence of SEQ ID        NO:832, and a light chain variable region comprising an amino        acid sequence that has at least 90% (e.g., at least 75%, 80%,        85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%)        sequence identity to SEQ ID NO:834 or that has the sequence of        SEQ ID NO:834; or    -   (n) a heavy chain variable region comprising an amino acid        sequence that has at least 90% (e.g., at least 75%, 80%, 85%,        90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence        identity to SEQ ID NO:837 or that has the sequence of SEQ ID        NO:837, and a light chain variable region comprising an amino        acid sequence that has at least 90% (e.g., at least 75%, 80%,        85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%)        sequence identity to SEQ ID NO:841 or that has the sequence of        SEQ ID NO:841; or    -   (o) a heavy chain variable region comprising an amino acid        sequence that has at least 90% (e.g., at least 75%, 80%, 85%,        90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence        identity to SEQ ID NO:845 or that has the sequence of SEQ ID        NO:845, and a light chain variable region comprising an amino        acid sequence that has at least 90% (e.g., at least 75%, 80%,        85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%)        sequence identity to SEQ ID NO:849 or that has the sequence of        SEQ ID NO:849; or    -   (p) a heavy chain variable region comprising an amino acid        sequence that has at least 90% (e.g., at least 75%, 80%, 85%,        90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence        identity to SEQ ID NO:852 or that has the sequence of SEQ ID        NO:852, and a light chain variable region comprising an amino        acid sequence that has at least 90% (e.g., at least 75%, 80%,        85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%)        sequence identity to SEQ ID NO:856 or that has the sequence of        SEQ ID NO:856; or    -   (q) a heavy chain variable region comprising an amino acid        sequence that has at least 90% (e.g., at least 75%, 80%, 85%,        90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence        identity to SEQ ID NO:859 or that has the sequence of SEQ ID        NO:859, and a light chain variable region comprising an amino        acid sequence that has at least 90% (e.g., at least 75%, 80%,        85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%)        sequence identity to SEQ ID NO:863 or that has the sequence of        SEQ ID NO:863; or    -   (r) a heavy chain variable region comprising an amino acid        sequence that has at least 90% (e.g., at least 75%, 80%, 85%,        90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence        identity to SEQ ID NO:867 or that has the sequence of SEQ ID        NO:867, and a light chain variable region comprising an amino        acid sequence that has at least 90% (e.g., at least 75%, 80%,        85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%)        sequence identity to SEQ ID NO:870 or that has the sequence of        SEQ ID NO:870; or    -   (s) a heavy chain variable region comprising an amino acid        sequence that has at least 90% (e.g., at least 75%, 80%, 85%,        90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence        identity to SEQ ID NO:873 or that has the sequence of SEQ ID        NO:873, and a light chain variable region comprising an amino        acid sequence that has at least 90% (e.g., at least 75%, 80%,        85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%)        sequence identity to SEQ ID NO:877 or that has the sequence of        SEQ ID NO:877; or    -   (t) a heavy chain variable region comprising an amino acid        sequence that has at least 90% (e.g., at least 75%, 80%, 85%,        90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence        identity to SEQ ID NO:880 or that has the sequence of SEQ ID        NO:880, and a light chain variable region comprising an amino        acid sequence that has at least 90% (e.g., at least 75%, 80%,        85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%)        sequence identity to SEQ ID NO:884 or that has the sequence of        SEQ ID NO:884; or    -   (u) a heavy chain variable region comprising an amino acid        sequence that has at least 90% (e.g., at least 75%, 80%, 85%,        90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence        identity to SEQ ID NO:888 or that has the sequence of SEQ ID        NO:888, and a light chain variable region comprising an amino        acid sequence that has at least 90% (e.g., at least 75%, 80%,        85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%)        sequence identity to SEQ ID NO:892 or that has the sequence of        SEQ ID NO:892; or    -   (v) a heavy chain variable region comprising an amino acid        sequence that has at least 90% (e.g., at least 75%, 80%, 85%,        90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence        identity to SEQ ID NO:894 or that has the sequence of SEQ ID        NO:894, and a light chain variable region comprising an amino        acid sequence that has at least 90% (e.g., at least 75%, 80%,        85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%)        sequence identity to SEQ ID NO:898 or that has the sequence of        SEQ ID NO:898; or    -   (w) a heavy chain variable region comprising an amino acid        sequence that has at least 90% (e.g., at least 75%, 80%, 85%,        90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence        identity to SEQ ID NO:902 or that has the sequence of SEQ ID        NO:902, and a light chain variable region comprising an amino        acid sequence that has at least 90% (e.g., at least 75%, 80%,        85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%)        sequence identity to SEQ ID NO:906 or that has the sequence of        SEQ ID NO:906; or    -   (x) a heavy chain variable region comprising an amino acid        sequence that has at least 90% (e.g., at least 75%, 80%, 85%,        90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence        identity to SEQ ID NO:910 or that has the sequence of SEQ ID        NO:910, and a light chain variable region comprising an amino        acid sequence that has at least 90% (e.g., at least 75%, 80%,        85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%)        sequence identity to SEQ ID NO:914 or that has the sequence of        SEQ ID NO:914; or    -   (y) a heavy chain variable region comprising an amino acid        sequence that has at least 90% (e.g., at least 75%, 80%, 85%,        90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence        identity to SEQ ID NO:917 or that has the sequence of SEQ ID        NO:917, and a light chain variable region comprising an amino        acid sequence that has at least 90% (e.g., at least 75%, 80%,        85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%)        sequence identity to SEQ ID NO:921 or that has the sequence of        SEQ ID NO:921; or    -   (z) a heavy chain variable region comprising an amino acid        sequence that has at least 90% (e.g., at least 75%, 80%, 85%,        90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence        identity to SEQ ID NO:925 or that has the sequence of SEQ ID        NO:925, and a light chain variable region comprising an amino        acid sequence that has at least 90% (e.g., at least 75%, 80%,        85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%)        sequence identity to SEQ ID NO:929 or that has the sequence of        SEQ ID NO:929.

Clone PA01P2D09

In some embodiments, a monoclonal antibody that specifically binds to amilk allergen comprises a heavy chain CDR1 comprising the amino acidsequence of SEQ ID NO:860, a heavy chain CDR2 comprising the amino acidsequence of SEQ ID NO:861, a heavy chain CDR3 comprising the amino acidsequence of SEQ ID NO:862, a light chain CDR1 comprising the amino acidsequence of SEQ ID NO:864, a light chain CDR2 comprising the amino acidsequence of SEQ ID NO:865, and a light chain CDR3 comprising the aminoacid sequence of SEQ ID NO:866.

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:859, and that comprises a        heavy chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:860, 861, and 862, respectively; and    -   (b) a light chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:863, and that comprises        the light chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:864, 865, and 866, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:859, and comprises a light chainvariable region comprising an amino acid sequence that has at least 90%sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, or 99% sequence identity) to SEQ ID NO:863. In some embodiments,the antibody comprises a heavy chain variable region comprising theamino acid sequence of SEQ ID NO:859 and comprises a light chainvariable region comprising the amino acid sequence of SEQ ID NO:863.

In some embodiments, the antibody is an antibody that competes forbinding with an antibody as described herein (e.g., an antibodycomprising a heavy chain CDR1-3 and a light chain CDR1-3 comprising theamino acid sequences of SEQ ID NOs:860, 861, 862, 864, 865, and 866,respectively, or an antibody comprising a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO:859 and furthercomprising a light chain variable region comprising the amino acidsequence of SEQ ID NO:863).

Clone PA01P2D04

In some embodiments, a monoclonal antibody that specifically binds to amilk allergen comprises a heavy chain CDR1 comprising the amino acidsequence of SEQ ID NO:121, a heavy chain CDR2 comprising the amino acidsequence of SEQ ID NO:826, a heavy chain CDR3 comprising the amino acidsequence of SEQ ID NO:827, a light chain CDR1 comprising the amino acidsequence of SEQ ID NO:829, a light chain CDR2 comprising the amino acidsequence of SEQ ID NO:830, and a light chain CDR3 comprising the aminoacid sequence of SEQ ID NO:831.

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:825, and that comprises a        heavy chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:121, 826, and 827, respectively; and    -   (b) a light chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:828, and that comprises        the light chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:829, 830, and 831, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:825, and comprises a light chainvariable region comprising an amino acid sequence that has at least 90%sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, or 99% sequence identity) to SEQ ID NO:828. In some embodiments,the antibody comprises a heavy chain variable region comprising theamino acid sequence of SEQ ID NO:825 and comprises a light chainvariable region comprising the amino acid sequence of SEQ ID NO:828.

In some embodiments, the antibody is an antibody that competes forbinding with an antibody as described herein (e.g., an antibodycomprising a heavy chain CDR1-3 and a light chain CDR1-3 comprising theamino acid sequences of SEQ ID NOs:121, 826, 827, 829, 830, and 831,respectively, or an antibody comprising a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO:825 and furthercomprising a light chain variable region comprising the amino acidsequence of SEQ ID NO:828).

Clone PA01P2B12

In some embodiments, a monoclonal antibody that specifically binds to amilk allergen comprises a heavy chain CDR1 comprising the amino acidsequence of SEQ ID NO:833, a heavy chain CDR2 comprising the amino acidsequence of SEQ ID NO:826, a heavy chain CDR3 comprising the amino acidsequence of SEQ ID NO:827, a light chain CDR1 comprising the amino acidsequence of SEQ ID NO:835, a light chain CDR2 comprising the amino acidsequence of SEQ ID NO:149, and a light chain CDR3 comprising the aminoacid sequence of SEQ ID NO:836.

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:832, and that comprises a        heavy chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:833, 826, and 827, respectively; and    -   (b) a light chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:834, and that comprises        the light chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:835, 149, and 836, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:832, and comprises a light chainvariable region comprising an amino acid sequence that has at least 90%sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, or 99% sequence identity) to SEQ ID NO:834. In some embodiments,the antibody comprises a heavy chain variable region comprising theamino acid sequence of SEQ ID NO:832 and comprises a light chainvariable region comprising the amino acid sequence of SEQ ID NO:834.

In some embodiments, the antibody is an antibody that competes forbinding with an antibody as described herein (e.g., an antibodycomprising a heavy chain CDR1-3 and a light chain CDR1-3 comprising theamino acid sequences of SEQ ID NOs:833, 826, 827, 835, 149, and 836,respectively, or an antibody comprising a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO:832 and furthercomprising a light chain variable region comprising the amino acidsequence of SEQ ID NO:834).

Clone PA01P2B05

In some embodiments, a monoclonal antibody that specifically binds to amilk allergen comprises a heavy chain CDR1 comprising the amino acidsequence of SEQ ID NO:868, a heavy chain CDR2 comprising the amino acidsequence of SEQ ID NO:378, a heavy chain CDR3 comprising the amino acidsequence of SEQ ID NO:869, a light chain CDR1 comprising the amino acidsequence of SEQ ID NO:871, a light chain CDR2 comprising the amino acidsequence of SEQ ID NO:682, and a light chain CDR3 comprising the aminoacid sequence of SEQ ID NO:872.

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:867, and that comprises a        heavy chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:868, 378, and 869, respectively; and    -   (b) a light chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:870, and that comprises        the light chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:871, 682, and 872, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:867, and comprises a light chainvariable region comprising an amino acid sequence that has at least 90%sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, or 99% sequence identity) to SEQ ID NO:870. In some embodiments,the antibody comprises a heavy chain variable region comprising theamino acid sequence of SEQ ID NO:867 and comprises a light chainvariable region comprising the amino acid sequence of SEQ ID NO:870.

In some embodiments, the antibody is an antibody that competes forbinding with an antibody as described herein (e.g., an antibodycomprising a heavy chain CDR1-3 and a light chain CDR1-3 comprising theamino acid sequences of SEQ ID NOs:868, 378, 869, 871, 682, and 872,respectively, or an antibody comprising a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO:867 and furthercomprising a light chain variable region comprising the amino acidsequence of SEQ ID NO:870).

Clone PA01P2D10

In some embodiments, a monoclonal antibody that specifically binds to amilk allergen comprises a heavy chain CDR1 comprising the amino acidsequence of SEQ ID NO:853, a heavy chain CDR2 comprising the amino acidsequence of SEQ ID NO:854, a heavy chain CDR3 comprising the amino acidsequence of SEQ ID NO:855, a light chain CDR1 comprising the amino acidsequence of SEQ ID NO:857, a light chain CDR2 comprising the amino acidsequence of SEQ ID NO:662, and a light chain CDR3 comprising the aminoacid sequence of SEQ ID NO:858.

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:852, and that comprises a        heavy chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:853, 854, and 855, respectively; and    -   (b) a light chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:856, and that comprises        the light chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:857, 662, and 858, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:852, and comprises a light chainvariable region comprising an amino acid sequence that has at least 90%sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, or 99% sequence identity) to SEQ ID NO:856. In some embodiments,the antibody comprises a heavy chain variable region comprising theamino acid sequence of SEQ ID NO:852 and comprises a light chainvariable region comprising the amino acid sequence of SEQ ID NO:856.

In some embodiments, the antibody is an antibody that competes forbinding with an antibody as described herein (e.g., an antibodycomprising a heavy chain CDR1-3 and a light chain CDR1-3 comprising theamino acid sequences of SEQ ID NOs:853, 854, 855, 857, 662, and 858,respectively, or an antibody comprising a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO:852 and furthercomprising a light chain variable region comprising the amino acidsequence of SEQ ID NO:856).

Clone PA01P2E08

In some embodiments, a monoclonal antibody that specifically binds to amilk allergen comprises a heavy chain CDR1 comprising the amino acidsequence of SEQ ID NO:800, a heavy chain CDR2 comprising the amino acidsequence of SEQ ID NO:801, a heavy chain CDR3 comprising the amino acidsequence of SEQ ID NO:802, a light chain CDR1 comprising the amino acidsequence of SEQ ID NO:804, a light chain CDR2 comprising the amino acidsequence of SEQ ID NO:110, and a light chain CDR3 comprising the aminoacid sequence of SEQ ID NO:805.

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:799, and that comprises a        heavy chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:800, 801, and 802, respectively; and    -   (b) a light chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:803, and that comprises        the light chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:804, 110, and 805, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:799, and comprises a light chainvariable region comprising an amino acid sequence that has at least 90%sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, or 99% sequence identity) to SEQ ID NO:803. In some embodiments,the antibody comprises a heavy chain variable region comprising theamino acid sequence of SEQ ID NO:799 and comprises a light chainvariable region comprising the amino acid sequence of SEQ ID NO:803

In some embodiments, the antibody is an antibody that competes forbinding with an antibody as described herein (e.g., an antibodycomprising a heavy chain CDR1-3 and a light chain CDR1-3 comprising theamino acid sequences of SEQ ID NOs:800, 801, 802, 804, 110, and 805,respectively, or an antibody comprising a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO:799 and furthercomprising a light chain variable region comprising the amino acidsequence of SEQ ID NO:803).

Antibodies that Specifically Bind to Fungal Allergens

In some embodiments, the monoclonal antibody or antigen-binding portionthereof specifically binds to a fungal allergen. In some embodiments,the monoclonal antibody specifically binds to a fungal allergen that isan Aspergillus fumigatus, Aspergillus niger, or Aspergillus nidulansallergen (e.g., an extract of Aspergillus fumigatus, Aspergillus niger,or Aspergillus nidulans). In some embodiments, the fungal allergen isAspergillus fumigatus 1 (Asp f 1), e.g., a purified recombinant allergenAspergillus fumigatus 1 (rAsp f 1).

In some embodiments, the monoclonal antibody exhibits cross-reactivitywith at least two fungal allergens. In some embodiments, the monoclonalantibody exhibits cross-reactivity with two or more Aspergillusallergens (e.g., two or more species of Aspergillus). In someembodiments, the monoclonal antibody exhibits cross-reactivity with twoor more of the fungal allergens Aspergillus fumigatus, Aspergillusniger, and Aspergillus nidulans. In some embodiments, the monoclonalantibody specifically binds to at least one of the fungal allergens witha K_(D) of less than 100 nM, less than 50 nM, less than 10 nM, less than5 nM, less than 1 nM, less than 500 pM, less than 250 pM, less than 150pM, less than 100 pM, less than 50 pM, less than 40 pM, less than 30 pM,less than 20 pM, less than about 10 pM, or less than 1 pM.

Antibody Sequences

In some embodiments, a monoclonal antibody that specifically binds to afungal allergen (e.g., that specifically binds to an Aspergillusallergen) comprises heavy chain CDRs and/or light chain CDRs that aredisclosed in Table 1 below. In some embodiments, a monoclonal antibodythat specifically binds to a fungal allerge (e.g., an Aspergillusallergen) comprises a light chain variable region sequence and/or aheavy chain variable region sequence sequence that is disclosed in Table1 below. In some embodiments, a monoclonal antibody that specificallybinds to a fungal allergen (e.g., an Aspergillus allergen) comprises: aheavy chain variable region comprising an amino acid sequence that hasat least 75% sequence identity (e.g., at least 80%, 85%, 90%, 91%, 92%,93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to a heavy chainvariable region sequence disclosed in Table 1 and comprises a CDR1, aCDR2, and a CDR3 that is identical to the CDRs of that heavy chainvariable region sequence, and a light chain variable region comprisingan amino acid sequence that has at least 70% sequence identity (e.g., atleast 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity to a light chain variable region sequence disclosed inTable 1 and comprises a CDR1, a CDR2, and a CDR3 that is identical tothe CDRs of that light chain variable region sequence.

In some embodiments, a monoclonal antibody that specifically binds to afungal allergen (e.g., that specifically binds to an Aspergillusallergen) comprises a light chain sequence, or a portion thereof, and/ora heavy chain sequence, or a portion thereof, derived from any of thefollowing antibodies described herein: Clone 1003320101_D6, Clone1003320105_D6, Clone 1003320107_C5, Clone 1003320107_F3, or Clone1003320107_F8. The amino acid sequences of the CDR, light chain variabledomain (VL), and heavy chain variable domain (VH) of Clone1003320101_D6, Clone 1003320105_D6, Clone 1003320107_C5, Clone1003320107_F3, and Clone 1003320107_F8 are set forth in Table 1 below.

In some embodiments, a monoclonal antibody that specifically binds to afungal allergen comprises one or more (e.g., one, two, three, four,five, or all six) of:

-   -   (a) a heavy chain CDR1 comprising the amino acid sequence of any        one of SEQ ID NOs:710, 718, 726, 734, or 742;    -   a heavy chain CDR2 comprising the amino acid sequence of any one        of SEQ ID NOs:711, 719, 727, 735, or 743;    -   a heavy chain CDR3 comprising the amino acid sequence of any one        of SEQ ID NOs:712, 720, 728, 736, or 744;    -   a light chain CDR1 comprising the amino acid sequence of any one        of SEQ ID NOs:714, 722, 730, 738, or 746;    -   a light chain CDR2 comprising the amino acid sequence of any one        of SEQ ID NOs:715, 723, 731, 739, or 747; and    -   a light chain CDR3 comprising the amino acid sequence of any one        of SEQ ID NOs:716, 724, 732, 740, or 748.

In some embodiments, a monoclonal antibody that specifically binds to afungal allergen comprises a heavy chain CDR1-3 and a light chain CDR1-3comprising the amino acid sequences of:

-   -   (a) SEQ ID NOs: 710, 711, 712, 714, 715, and 716, respectively;        or    -   (b) SEQ ID NOs:718, 719, 720, 722, 723, and 724, respectively;        or    -   (c) SEQ ID NOs:726, 7272, 728, 730, 731, and 732, respectively;        or    -   (d) SEQ ID NOs:734, 735, 736, 738, 739, and 740, respectively;        or    -   (e) SEQ ID NOs:742, 743, 744, 746, 747, and 748, respectively.

In some embodiments, a monoclonal antibody that specifically binds to afungal allergen comprises a heavy chain variable region comprising anamino acid sequence that has at least 75% sequence identity (e.g., atleast 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity to any one of SEQ ID NOs:709, 717, 725, 733, or 741.In some embodiments, a monoclonal antibody that specifically binds to afungal allergen comprises a light chain variable region comprising anamino acid sequence that has at least 75% sequence identity (e.g., atleast 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity to any one of SEQ ID NOs:713, 721, 729, 737, or 745.In some embodiments, a monoclonal antibody that specifically binds to afungal allergen comprises a heavy chain variable region comprising anamino acid sequence that has at least 75% sequence identity (e.g., atleast 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity to any one of SEQ ID NOs:709, 717, 725, 733, or 741,and comprises a light chain variable region comprising an amino acidsequence that has at least 75% sequence identity (e.g., at least 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequenceidentity to any one of SEQ ID NOs:713, 721, 729, 737, or 745.

In some embodiments, a monoclonal antibody that specifically binds to afungal allergen comprises a heavy chain variable region comprising anamino acid sequence that has at least 75% sequence identity (e.g., atleast 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity to any one of SEQ ID NOs:709, 717, 725, 733, or 741and comprises a CDR1, a CDR2, and a CDR3 that is identical to the CDRsof that SEQ ID NO. In some embodiments, a monoclonal antibody thatspecifically binds to a fungal allergen comprises a light chain variableregion comprising an amino acid sequence that has at least 70% sequenceidentity (e.g., at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%,96%, 97%, 98%, or 99% sequence identity to any one of SEQ ID NOs:713,721, 729, 737, or 745 and comprises a CDR1, a CDR2, and a CDR3 that isidentical to the CDRs of that SEQ ID NO. In some embodiments, amonoclonal antibody that specifically binds to a fungal allergencomprises:

-   -   (a) a heavy chain variable region comprising an amino acid        sequence that has at least 75% sequence identity (e.g., at least        80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%        sequence identity to any one of SEQ ID NOs:709, 717, 725, 733,        or 741 and that comprises a CDR1, a CDR2, and a CDR3 that is        identical to the CDRs of that SEQ ID NO; and    -   (b) a light chain variable region comprising an amino acid        sequence that has at least 75% sequence identity (e.g., at least        80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%        sequence identity to any one of SEQ ID NOs:713, 721, 729, 737,        or 745 and that comprises a CDR1, a CDR2, and a CDR3 that is        identical to the CDRs of that SEQ ID NO.

Clone 1003320101 D6

In some embodiments, a monoclonal antibody that specifically binds to afungal allergen (e.g., Aspergillus allergen) comprises a heavy chainCDR1 comprising the amino acid sequence of SEQ ID NO:710, a heavy chainCDR2 comprising the amino acid sequence of SEQ ID NO:711, a heavy chainCDR3 comprising the amino acid sequence of SEQ ID NO:712, a light chainCDR1 comprising the amino acid sequence of SEQ ID NO:714, a light chainCDR2 comprising the amino acid sequence of SEQ ID NO:715, and a lightchain CDR3 comprising the amino acid sequence of SEQ ID NO:716. In someembodiments, the antibody specifically binds to the fungal allergenAspergillus fumigatus.

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:709, and that comprises a        heavy chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:710, 711, and 712, respectively; and    -   (b) a light chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:713, and that comprises        the light chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:714, 715, and 716, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:709, and comprises a light chainvariable region comprising an amino acid sequence that has at least 90%sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, or 99% sequence identity) to SEQ ID NO:713. In some embodiments,the antibody comprises a heavy chain variable region comprising theamino acid sequence of SEQ ID NO:709 and comprises a light chainvariable region comprising the amino acid sequence of SEQ ID NO:713.

In some embodiments, the antibody is an antibody that competes forbinding with an antibody as described herein (e.g., an antibodycomprising a heavy chain CDR1-3 and a light chain CDR1-3 comprising theamino acid sequences of SEQ ID NOs:710, 711, 712, 714, 715, and 716,respectively, or an antibody comprising a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO:709 and furthercomprising a light chain variable region comprising the amino acidsequence of SEQ ID NO:713).

Clone 1003320105 D6

In some embodiments, a monoclonal antibody that specifically binds to afungal allergen (e.g., Aspergillus allergen) comprises a heavy chainCDR1 comprising the amino acid sequence of SEQ ID NO:718, a heavy chainCDR2 comprising the amino acid sequence of SEQ ID NO:719, a heavy chainCDR3 comprising the amino acid sequence of SEQ ID NO:720, a light chainCDR1 comprising the amino acid sequence of SEQ ID NO:722, a light chainCDR2 comprising the amino acid sequence of SEQ ID NO:723, and a lightchain CDR3 comprising the amino acid sequence of SEQ ID NO:724. In someembodiments, the antibody specifically binds to the fungal allergenAspergillus fumigatus.

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:717, and that comprises a        heavy chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:718, 719, and 720, respectively; and    -   (b) a light chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:721, and that comprises        the light chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:722, 723, and 724, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:717, and comprises a light chainvariable region comprising an amino acid sequence that has at least 90%sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, or 99% sequence identity) to SEQ ID NO:721. In some embodiments,the antibody comprises a heavy chain variable region comprising theamino acid sequence of SEQ ID NO:717 and comprises a light chainvariable region comprising the amino acid sequence of SEQ ID NO:721.

In some embodiments, the antibody is an antibody that competes forbinding with an antibody as described herein (e.g., an antibodycomprising a heavy chain CDR1-3 and a light chain CDR1-3 comprising theamino acid sequences of SEQ ID NOs:718, 719, 720, 722, 723, and 724,respectively, or an antibody comprising a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO:717 and furthercomprising a light chain variable region comprising the amino acidsequence of SEQ ID NO:721).

Clone 1003320107 C5

In some embodiments, a monoclonal antibody that specifically binds to afungal allergen (e.g., Aspergillus allergen) comprises a heavy chainCDR1 comprising the amino acid sequence of SEQ ID NO:726, a heavy chainCDR2 comprising the amino acid sequence of SEQ ID NO:727, a heavy chainCDR3 comprising the amino acid sequence of SEQ ID NO:728, a light chainCDR1 comprising the amino acid sequence of SEQ ID NO:730, a light chainCDR2 comprising the amino acid sequence of SEQ ID NO:731, and a lightchain CDR3 comprising the amino acid sequence of SEQ ID NO:732. In someembodiments, the antibody specifically binds to the fungal allergenAspergillus fumigatus. In some embodiments, the antibody specificallybinds to the fungal allergen Aspergillus niger. In some embodiments, theantibody specifically binds to the fungal allergen Aspergillus nidulans.In some embodiments, the antibody specifically binds to a recombinantAspergillus antigen (e.g., rAsp f 1). In some embodiments, the antibodyspecifically binds cross-reactively to more than one of Aspergillusfumigatus, Aspergillus niger, Aspergillus nidulans, or a recombinantAspergillus antigen (e.g., rAsp f 1).

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:725, and that comprises a        heavy chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:726, 727, and 728, respectively; and    -   (b) a light chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:729, and that comprises        the light chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:730, 731, and 732, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:725, and comprises a light chainvariable region comprising an amino acid sequence that has at least 90%sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, or 99% sequence identity) to SEQ ID NO:729. In some embodiments,the antibody comprises a heavy chain variable region comprising theamino acid sequence of SEQ ID NO:725 and comprises a light chainvariable region comprising the amino acid sequence of SEQ ID NO:729.

In some embodiments, the antibody is an antibody that competes forbinding with an antibody as described herein (e.g., an antibodycomprising a heavy chain CDR1-3 and a light chain CDR1-3 comprising theamino acid sequences of SEQ ID NOs:726, 727, 728, 730, 731, and 732,respectively, or an antibody comprising a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO:725 and furthercomprising a light chain variable region comprising the amino acidsequence of SEQ ID NO:729).

Clone 1003320107 F3

In some embodiments, a monoclonal antibody that specifically binds to afungal allergen (e.g., Aspergillus allergen) comprises a heavy chainCDR1 comprising the amino acid sequence of SEQ ID NO:734, a heavy chainCDR2 comprising the amino acid sequence of SEQ ID NO:735, a heavy chainCDR3 comprising the amino acid sequence of SEQ ID NO:736, a light chainCDR1 comprising the amino acid sequence of SEQ ID NO:738, a light chainCDR2 comprising the amino acid sequence of SEQ ID NO:739, and a lightchain CDR3 comprising the amino acid sequence of SEQ ID NO:740. In someembodiments, the antibody specifically binds to a recombinantAspergillus antigen (e.g., rAsp f 1).

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:733, and that comprises a        heavy chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:734, 735, and 736, respectively; and    -   (b) a light chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:737, and that comprises        the light chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:738, 739, and 740, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:733, and comprises a light chainvariable region comprising an amino acid sequence that has at least 90%sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, or 99% sequence identity) to SEQ ID NO:737. In some embodiments,the antibody comprises a heavy chain variable region comprising theamino acid sequence of SEQ ID NO:733 and comprises a light chainvariable region comprising the amino acid sequence of SEQ ID NO:737.

In some embodiments, the antibody is an antibody that competes forbinding with an antibody as described herein (e.g., an antibodycomprising a heavy chain CDR1-3 and a light chain CDR1-3 comprising theamino acid sequences of SEQ ID NOs:734, 735, 736, 738, 739, and 740,respectively, or an antibody comprising a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO:733 and furthercomprising a light chain variable region comprising the amino acidsequence of SEQ ID NO:737).

Clone 1003320107 F8

In some embodiments, a monoclonal antibody that specifically binds to afungal allergen (e.g., Aspergillus allergen) comprises a heavy chainCDR1 comprising the amino acid sequence of SEQ ID NO:742, a heavy chainCDR2 comprising the amino acid sequence of SEQ ID NO:743, a heavy chainCDR3 comprising the amino acid sequence of SEQ ID NO:744, a light chainCDR1 comprising the amino acid sequence of SEQ ID NO:746, a light chainCDR2 comprising the amino acid sequence of SEQ ID NO:747, and a lightchain CDR3 comprising the amino acid sequence of SEQ ID NO:748. In someembodiments, the antibody specifically binds to the fungal allergenAspergillus fumigatus.

In some embodiments, the antibody comprises:

-   -   (a) a heavy chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:741, and that comprises a        heavy chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:74222, 743, and 744, respectively; and    -   (b) a light chain variable region that comprises an amino acid        sequence that has at least 70% sequence identity (e.g., at least        70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,        or 99% sequence identity) to SEQ ID NO:745, and that comprises        the light chain CDR1, CDR2, and CDR3 comprising the amino acid        sequences of SEQ ID NOs:746, 747, and 748, respectively.

In some embodiments, the antibody comprises a heavy chain variableregion comprising an amino acid sequence that has at least 90% sequenceidentity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%sequence identity) to SEQ ID NO:741, and comprises a light chainvariable region comprising an amino acid sequence that has at least 90%sequence identity (e.g., at least 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, or 99% sequence identity) to SEQ ID NO:745. In some embodiments,the antibody comprises a heavy chain variable region comprising theamino acid sequence of SEQ ID NO:741 and comprises a light chainvariable region comprising the amino acid sequence of SEQ ID NO:745.

In some embodiments, the antibody is an antibody that competes forbinding with an antibody as described herein (e.g., an antibodycomprising a heavy chain CDR1-3 and a light chain CDR1-3 comprising theamino acid sequences of SEQ ID NOs:742, 743, 744, 746, 747, and 748,respectively, or an antibody comprising a heavy chain variable regioncomprising the amino acid sequence of SEQ ID NO:741 and furthercomprising a light chain variable region comprising the amino acidsequence of SEQ ID NO:745).

Antigen-Binding Fragments

In some embodiments, an antibody as disclosed herein (e.g., an antibodyas disclosed in Section IV that binds to a food allergen, plantallergen, fungal allergen, animal allergen, dust mite allergen, drugallergen, cosmetic allergen, or latex allergen) is an antigen-bindingportion (also referred to herein as an antigen-binding fragment).Examples of antigen-binding fragments include, but are not limited to, aFab, a F(ab′)₂, a Fv, a scFv, a bivalent scFv, a single domain antibody,or a diabody. Various techniques have been developed for the productionof antigen-binding fragments. Traditionally, these fragments werederived via proteolytic digestion of intact antibodies (see, e.g.,Morimoto et al., J. Biochem. Biophys. Meth., 24:107-117 (1992); andBrennan et al., Science, 229:81 (1985)). However, these fragments cannow be produced directly using recombinant host cells. For example,antigen-binding fragments can be isolated from antibody phage libraries.Alternatively, Fab′-SH fragments can be directly recovered from E. colicells and chemically coupled to form F(ab′)2 fragments (see, e.g.,Carter et al., BioTechnology, 10:163-167 (1992)). According to anotherapproach, F(ab′)₂ fragments can be isolated directly from recombinanthost cell culture. Other techniques for the production ofantigen-binding fragments are known in the art.

Antibody Conjugates

In some embodiments, the antibody or antigen-binding fragment can beconjugated to another molecule, e.g., polyethylene glycol (PEGylation)or serum albumin, to provide an extended half-life in vivo. Examples ofPEGylation of antigen-binding fragments are provided in Knight et al.Platelets 15:409, 2004 (for abciximab); Pedley et al., Br. J. Cancer70:1126, 1994 (for an anti-CEA antibody); Chapman et al., NatureBiotech. 17:780, 1999; and Humphreys, et al., Protein Eng. Des. 20: 227,2007).

In some embodiments, antibody-drug conjugates comprising a monoclonalantibody or antigen-binding fragment as described herein are provided.In some embodiments, a monoclonal antibody or antigen-binding fragment(e.g., an antibody or antigen-binding fragment that specifically bindsto a food allergen or a fungal allergen) is covalently linked to acytotoxic drug. In some embodiments, the antibody or antigen-bindingfragment is an antibody that specifically binds to a fungal allergen andthe drug is an anti-fungal drug. Suitable anti-fungal drugs include, butare not limited to, Amphotericin B, azole anti-fungals (e.g.,ketoconazole, fluconazole, isavuconazole, itraconazole, posaconazole, orvoriconazole), echinocandins (e.g., anidulafungin, caspofungin, ormicafungin), and flucytosine. Methods for making antibody-drugconjugates are described, e.g., in Chudasama et al., Nature Chemistry,2016, 8:114-119; WO 2013/068874; and U.S. Pat. No. 8,535,678.

Synthetic Antibodies, Antibody Compositions, and Antibody-ProducingCells

Certain antibodies described herein are derived from B cells isolatedfrom human subjects who have been exposed to allergen(s). In certainembodiments, antibodies, antibody compositions, and cells of theinvention are distinguishable from naturally occurring antibodies,compositions and cells in one or more respects. The distinguishableantibodies, compositions, and cells may be referred to as “synthetic,”or may be identified by the proviso that the antibody or composition “isnot naturally occurring” or affirmatively as “non-naturally occurring.”As used herein the terms “corresponding antibody,” and “correspondingto” describes the relationship between (1) an antibody characterized bysix specific CDR sequences and produced by immune cells of a studysubject described in the Examples below and (2) a synthetic antibodycomprising the same six CDR sequences.

Synthetic Antibodies

Synthetic antibodies of the invention may differ in structure fromnaturally occurring antibodies with the same CDRs. That is, syntheticantibodies identified by specified CDRs may be structurally differentfrom antibodies comprising the specified CDRs that are produced by cellsof the study subject described in the Examples below. Possibledifferences include:

Variable Region Sequences that Differ Corresponding Naturally OccurringAntibodies

In one approach, an antibody heavy chain comprises the CDRs of a clonedescribed herein (e.g., PA13P1E10) with the proviso that the antibodyheavy chain does not comprise the heavy chain variable region sequenceassociated with the clone described herein. For illustration, in oneembodiment an antibody that comprises the CDRs of Clone PA13P1E10 doesnot have a heavy chain variable region that comprises SEQ ID NO:9. Inanother approach, an antibody light chain comprises the CDRs of a clonedescribed herein (e.g., PA13P1E10) with the proviso that the antibodylight chain does not comprise the light chain variable region sequenceassociated with the clone described herein. For illustration, in oneembodiment an antibody that comprises the CDRs of Clone PA13P1E10 doesnot have a light chain variable region that comprises SEQ ID NO:13). Inone approach both the heavy chain and the light chain variable region ofan antibody of the invention have an amino acid sequence other than thesequence disclosed herein.

Lambda and Kappa Light Chains

In some embodiments the synthetic antibody comprises lambda type lightchains. In some embodiments the synthetic antibody comprises kappa typelight chains.

Isotypes

In some embodiments the synthetic antibody with specified CDRs is anisotype other the isotype(s) found associated with the study subjectfrom which B cells with the specified CDRs was derived. In someembodiments the antibody disclosed herein is an isotype other than IgG1.In some embodiments the antibody disclosed herein is an isotype otherthan IgG2. In some embodiments the antibody disclosed herein is anisotype other than IgG3. In some embodiments the antibody disclosedherein is an isotype other than IgG4. In some embodiments the antibodydisclosed herein is an isotype other than IgM. In some embodiments theantibody disclosed herein is an isotype other than IgA.

Allotypes

In some embodiments the synthetic antibody with specified CDRs is anallotype other the allotype(s) found associated with the study subjectfrom which B cells with the specified CDRs was derived. In someembodiments, the synthetic antibody of the invention comprises anallotype selected from those listed in Table 2, below, which isdifferent from an allotype of antibodies from the corresponding studysubject. In some embodiments the synthetic antibody of the inventioncomprises any individual allotype selected from those listed in Table 2,with the proviso that the allotype differs from the correspondingallotype of antibodies from a study subject.

TABLE 2 Human immunoglobulin allotypes Heavy chains Isotype/type IgG1IgG2 IgG3 IgA Light chains Allotypes G1m G2m G3m A2m Km 1 (a) 23 (n) 21(g1) 1 1 2 (x) 28 (g5) 2 2 3 (f) 11 (b0) 3 17 (z) 5 (b1) 13 (b3) 14 (b4)10 (b5) 15 (s) 16 (t) 6 (c3) 24 (c5) 26 (u) 27 (v) NB: Alphabeticalnotation given within brackets. From: Jefferis and Marie-Paule Lefranc,2009, “Human immunoglobulin allotypes: Possible implications forimmunogenicity” mAbs 1(4): 332-338, incorporated herein by reference.

Constant Domain Variants

Synthetic antibodies of the invention may comprise variations in heavychain constant regions to change the properties of the syntheticantibody relative to the corresponding naturally occurring antibody.Exemplary changes include mutations to modulate antibody effectorfunction (e.g., complement-based effector function or FcγR-basedeffector function), alter half-like, modulate coengagement of antigenand FcγRs, introduce or remove glycosylation motifs (glyco-engineering).See Fonseca et al., 2018, “Boosting half-life and effector functions oftherapeutic antibodies by Fc-engineering: An interaction-functionreview” Intl Biol Macromol. 19:306-311; Wang et al., 2018, “IgG Fcengineering to modulate antibody effector functions” Protein Cell 2018,9(1):63-73; Schlothauer, 2016, “Novel human IgG1 and IgG4 Fc-engineeredantibodies with completely abolished immune effector functions,” ProteinEngineering, Design and Selection 29(10):457-466; Tam et al., 2017,“Functional, Biophysical, and Structural Characterization of Human IgG1and IgG4 Fc Variants with Ablated Immune Functionality” Antibodies 6,12, each incorporated herein by reference for all purposes.

Synthetic Antibody Compositions

Synthetic antibody compositions of the invention may differ fromnaturally occurring compositions in at least one or more of thefollowing respects: (i) composition comprises antibodies that arepurified, i.e., separated from tissue or cellular material with whichthey are associated in the human body, and optionally in an manufacturedexcipient or medium; and/or (ii) antibody compositions of the inventioncontain a single species of antibody (are monoclonal) such that allantibodies in the composition have the same structure and specificity;

Synthetic Antibody-Producing Cells

Antibodies described herein may be produced by recombinant expression ina human or non-human cell. Synthetic antibody-producing cells includenon-human cells expressing heavy chains, light chains, or both heavy andlight chains; human cells that are not immune cells heavy chains, lightchains, or both heavy and light chains; and human B cells that produceheavy chains or light chains, but not both heavy and light chains.Synthetic antibodies of the invention may be are heterologouslyexpressed, in vitro or in vivo, in cells other than human B cells, suchas non-human cells and human cells other than B cells, optionally otherthan immune cells, and optionally in cells other than cells in a B celllineage.

V. Methods of Use

In another aspect, the present disclosure provides therapeutic methodsfor treating a human subject with one or more of the allergen-specificmonoclonal antibodies or antigen-binding portions thereof as disclosedherein. In some embodiments, methods of treating an allergy areprovided. In some embodiments, methods of reducing one or more allergysymptoms in a subject are provided. In some embodiments, theallergen-specific monoclonal antibodies disclosed herein are usedtherapeutically as blocking antibodies, which is often referred to aspassive immunotherapy. Without being bound to a particular theory, it ishypothesized that the allergen-specific monoclonal antibodies disclosedherein block allergen binding to IgE or outcompete endogenous IgE forallergen binding, which in turns prevents or reduces initiation of theallergic cascade. Without intending to be bound by a particularmechanism in some embodiments antibodies of the invention providetherapeutic benefit by binding inhibitory receptors on mast cells and/orbasophils.

In some embodiments, the method comprises administering to the subject atherapeutically effective amount of one or more allergen-specificmonoclonal antibodies as disclosed herein (e.g., one or moreallergen-specific monoclonal antibodies as disclosed in Section IVabove). In some embodiments, the method comprises administering to thesubject a therapeutically effective amount of a pharmaceuticalcomposition comprising one or more allergen-specific monoclonalantibodies as disclosed herein (e.g., a pharmaceutical composition asdisclosed in Section VI below).

In some embodiments, the method comprises administering to the subject atherapeutically effective amount of an allergen-specific monoclonalantibody that is a human IgG isotype, such as a human IgG4 isotype, orantigen-binding portion thereof comprising at least a portion of a humanIgG or IgG4 isotype constant region sequence.

In some embodiments, the method comprises administering to the subject atherapeutically effective amount of an allergen-specific monoclonalantibody or antigen-binding portion thereof. In some embodiments, themethod comprises administering to the subject two or moreallergen-specific monoclonal antibodies (e.g., in a pharmaceuticalcomposition comprising the two or more allergen-specific monoclonalantibodies). In some embodiments, the method comprises administering twoor more antibodies that specifically bind to the same allergen. In someembodiments, the method comprises administering two or more antibodiesthat specifically bind to different epitopes of the same allergen. Insome embodiments, the method comprises administering two or moreantibodies that specifically bind to two or more different allergens.

In some embodiments, the therapeutic antibody is an antibody thatcomprises CDR sequences, a heavy chain variable region, and/or a lightchain variable region as described herein (e.g., as disclosed in Table 1below) and further comprises a native or modified IgM, IgD, IgG3, IgG1,IgA1, IgG2, IgG4, or IgA2 heavy chain constant region.

In some embodiments, the therapeutic antibody is conjugated to a drug,e.g., as described in Section IV above.

In some embodiments, the human subject to be treated is an adult. Insome embodiments, the human subject is a juvenile.

In some embodiments, a human subject to be treated has an allergy to afood allergen, a plant allergen, a fungal allergen, an animal allergen,a dust mite allergen, a drug allergen, a cosmetic allergen, or a latexallergen. In some embodiments, the human subject has an allergy to afood allergen. In some embodiments, the food allergen is a milkallergen, an egg allergen, a nut allergen, a fish allergen, a shellfishallergen, a soy allergen, a legume allergen, a seed allergen, or a wheatallergen. In some embodiments, the food allergen is a peanut allergen.In some embodiments, the food allergen is a milk allergen. In someembodiments, the food allergen is an egg allergen. In some embodiments,the human subject has an allergy to a plant allergen or a fungalallergen (e.g., an Aspergillus allergen). In some embodiments, theallergen is a pollen allergen (e.g., tree pollen, grass pollen, or weedpollen) or a mold allergen. In some embodiments, the human subject hasan allergy to an animal allergen. In some embodiments, the allergen is adander allergen or an insect sting.

In some embodiments, the human subject to be treated has allergies totwo or more allergens, e.g., to two or more of a food allergen, a plantallergen, a fungal allergen, an animal allergen, a dust mite allergen, adrug allergen, a cosmetic allergen, or a latex allergen. In someembodiments, the human subject has allergies to 2, 3, 4, 5, 6, 7, 8, 9,10 or more allergens. In some embodiments, the human subject hasallergies to two or more different types of antigens (allergens) in aclass of allergen, e.g., allergies to two or more different foodallergens (e.g., allergies to two or more different peanut antigens, orallergies to a peanut allergen and a non-peanut allergen such as an eggor milk allergen). In some embodiments, the human subject has allergiesto two more different classes of allergens (e.g., allergies to one ormore food allergens and to one or more plant allergens). In someembodiments, a human subject has an allergy to 1, 2, 3, 4, 5, 6, 7, 8,9, 10 or more allergens in the same class of allergen but does not haveany known allergies to allergens in other classes of allergens. Forexample, in some embodiments, a human subject has an allergy to 1, 2, 3,4, 5, 6, 7, 8, 9, 10 or more food allergens but does not have any knownallergies to non-food allergens. In some embodiments, a human subjecthas an allergy to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more fungal allergensbut does not have any known allergies to non-fungal allergens.

In some embodiments, the therapeutic methods disclosed herein reduce oneor more symptoms of the allergy in the subject. It will be appreciatedby a person of ordinary skill in the art that the symptom(s) associatedwith an allergic reaction can vary depending upon the type of allergenthat induces the allergic reaction. Examples of allergic reactionsymptoms include, but are not limited to, hives, rashes, eczema flare,redness of skin, itchy mouth, itchy eyes, nausea, vomiting, diarrhea,stomach pain, nasal congestion, runny nose, stuffy nose, sneezing,cough, fatigue, sore throat, swelling of the lips, tongue, or throat,headaches, trouble swallowing, shortness of breath, wheezing, drop inblood pressure, or weak pulse. In some embodiments, the therapeuticmethods disclosed herein reduce the severity of one or more symptoms ofthe allergy. In some embodiments of the therapeutic methods describedherein, the allergy symptoms in the subject comprise one or more ofrunny nose, skin hives, skin redness, skin swelling, itching or tinglingin or around the mouth and/or throat, difficulty swallowing, wateryeyes, diarrhea, stomach cramps, nausea, vomiting, tightening of thethroat, shortness of breath or wheezing, shortness of breath, andanaphylaxis. In some embodiments, the therapeutic methods disclosedherein reduce the length of duration of one or more symptoms of theallergy.

In some embodiments, the therapeutic methods disclosed herein reduce oneor more symptoms of allergic reaction to an allergen such as a foodallergen (e.g., a peanut allergen), such as but not limited to hives,rashes, eczema flare, redness of skin, itchy mouth, nausea, vomiting,diarrhea, stomach pain, nasal congestion, runny nose, sneezing, drycough, swelling of the lips, tongue, or throat, trouble swallowing,shortness of breath, wheezing, drop in blood pressure, or weak pulse. Insome embodiments, administration of one or more allergen-specificmonoclonal antibodies as disclosed herein reduces the severity of one ormore of the symptoms and/or reduces the length of duration of one ormore of the symptoms.

In some embodiments, an allergen-specific monoclonal antibody asdisclosed herein is administered to a human subject at a therapeuticallyeffective amount or dose. In some embodiments, a daily dose range ofabout 0.01 mg/kg to about 500 mg/kg, or about 0.1 mg/kg to about 200mg/kg, or about 1 mg/kg to about 100 mg/kg, or about 10 mg/kg to about50 mg/kg, can be used. The dosages, however, may be varied according toseveral factors, including the chosen route of administration, theformulation of the composition, patient response, the severity of thecondition, the subject's weight, and the judgment of the prescribingphysician. The dosage can be increased or decreased over time, asrequired by an individual patient. In certain instances, a patientinitially is given a low dose, which is then increased to an efficaciousdosage tolerable to the patient. Determination of an effective amount iswell within the capability of those skilled in the art.

The route of administration of an antibody or composition comprising anantibody as described herein can be dermal or transdermal, inhalational,intestinal, intravenous, intramuscular, intraperitoneal, intrathecal,intralesional, intrabronchial, nasal, ocular or otic delivery, oral,rectal, subcutaneous, topical, transmucosal, or any other methods knownin the art. In some embodiments, the antibody or composition isadministered by infusion (e.g., intravenously) or by injection (e.g.,subcutaneously). In some embodiments, the route of administration of anantibody or composition comprising an antibody in any of the methodsdescribed herein is subcutaneous, intravenous, or intranasal.

In some embodiments, administration of a single dose of an antibody orcomposition comprising an antibody as described herein is effective totreat the allergy or reduce one or more symptoms of the allergy. In someembodiments, multiple doses of the antibody or composition areadministered. In some embodiments, a second dose is administered atleast 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days or longer,e.g., at least 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7weeks, 8 weeks, or longer, after administration of the first dose. Insome embodiments, an antibody or composition comprising an antibody asdescribed herein is administered to a subject about every 1, 2, 3, 4, 5,6, 7, 8, 9, or 10 week(s). In some embodiments, an antibody orcomposition comprising an antibody as described herein is administeredto a subject over an extended period of time, e.g., for at least 30, 40,50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 350 days or longer.

In some embodiments, in any of the methods described herein, the subjectis further administered an additional agent, e.g., an antihistamine, anepinephrine, a decongestant, a bronchial dilator, or a corticosteroid.In some embodiments, the monoclonal antibody and the additional agentare administered substantially simultaneously, i.e., in the samepharmaceutical composition or in separation pharmaceutical compositionsthat are administered at substantially the same time (e.g., administeredwithin seconds of each other). In some embodiments, the monoclonalantibody and the additional agent are administered separately. In someembodiments, the monoclonal antibody is administered first, followed byadministering of the additional agent. In some embodiments, theadditional agent is administered first, followed by administering of themonoclonal antibody.

In some embodiments, in any of the methods described herein, the methodscan further comprise a step of assessing the reduction of the allergysymptoms (e.g., allergy symptoms related to a peanut allergy, a tree nutallergy, a milk allergy, or a fungal allergy) in the subject. In someembodiments, the reduction of the allergy symptoms can be measured by aTotal Nasal Symptom Score (TNSS), which is made from patient assessmentof four symptoms graded on a 0 (none) to 3 (severe) scale forcongestion, itching, and rhinorrhea, and 0 (none) to 3 (5 or moresneezes) for sneezing. Each of the four symptoms is evaluated using thefollowing scale of 0=None, 1=Mild, 2=Moderate, or 3=Severe. The TNSS hasa possible score of 0-12. In other embodiments, the reduction of theallergy symptoms can be measured by a Visual Analog Scale (VAS) nasalsymptoms score, which is often used to classify allergy burden intomild, moderate, and severe. A VAS nasal symptoms score ranging from 0(no nasal symptoms) to 100 (maximal nasal symptoms) can be used toassess the severity of combined nasal symptoms. In other embodiments,the reduction of the allergy symptoms can be measured by peak nasalinspiratory flow (PNIF), which uses a nasal spirometer to measure thenasal airflow (measured as 1/min) in a patient. In yet otherembodiments, the reduction of the allergy symptoms can be measured by anallergen skin test, such as a skin prick test (SPT), which uses thepresence and degree of cutaneous reactivity as a marker forsensitization within target organs, such as eyes, nose, lung, gut andskin. When relevant allergens (e.g., a peanut allergen, a tree nutallergen, a milk allergen, or a fungal allergen) are introduced into theskin, allergic reactions on the skin produce a wheal and flare responsethat can be quantitated, for example, using the diameter of the wheal.In yet other embodiments, the reduction of the allergy symptoms can bemeasured by basophil activation test, which utilizes flow cytometry toquantify the expression of markers of activation on the surface ofbasophils following allergen stimulation. In yet other embodiments, thereduction of the allergy symptoms can be measured by oral foodchallenge, which involves administering escalating doses of an allergento an allergic individual under the supervision of a trained allergistor immunologist. An oral food challenge may be conducted according to anopen, single-blind, or double-blind format, with the gold-standard beingboth double-blind and placebo-controlled.

In yet another aspect, the present disclosure provides diagnostic anddetection methods using one or more of the allergen-specific monoclonalantibodies or antigen-binding portions thereof as disclosed herein. Insome embodiments, an allergen-specific monoclonal antibody orantigen-binding portion thereof is used to detect whether a sample froma subject has allergic reactivity to an allergen (e.g., a food allergensuch as a peanut allergen, tree nut allergen, or milk allergen), a plantallergen, a fungal allergen, an animal allergen, a dust mite allergen, adrug allergen, a cosmetic allergen, or a latex allergen. In someembodiments, the allergen-specific monoclonal antibody orantigen-binding portion thereof is used to detect whether a sample froma subject has allergic reactivity to a specific epitope of the allergen(e.g., using an antibody that is known to bind to a specific epitope ofthe allergen). In some embodiments, the method comprises contacting asample from the subject (e.g., a blood or plasma sample) with anallergen-specific monoclonal antibody or antigen-binding portion asdisclosed herein.

VI. Compositions and Kits

In another aspect, compositions and kits comprising one or moreallergen-specific monoclonal antibodies or antigen-binding portionsthereof that are generated from human B cells are provided.

Pharmaceutical Compositions

In some embodiments, pharmaceutical compositions comprising one or moreallergen-specific monoclonal antibodies or antigen-binding portionsthereof are provided. In some embodiments, the pharmaceuticalcomposition comprises a monoclonal antibody as described herein, e.g.,as disclosed in Section IV above. In some embodiments, thepharmaceutical composition is for use in a method of reducing one ormore allergy symptoms in a subject (e.g., allergy symptoms due to anallergy to a food allergen, a plant allergen, a fungal allergen, ananimal allergen, a dust mite allergen, a drug allergen, a cosmeticallergen, or a latex allergen). In some embodiments, the pharmaceuticalcomposition is for use in a method of reducing one or more allergysymptoms in a subject having a food allergy, e.g., a peanut allergy. Insome embodiments, the pharmaceutical composition is for use in a methodof reducing one or more allergy symptoms in a subject having an allergyto two more allergens (e.g., two or more food allergens, e.g., peanutallergy and tree nut allergy). In some embodiments, the pharmaceuticalcomposition is for use in a method of reducing one or more allergysymptoms in a subject having a fungal allergy.

In some embodiments, the pharmaceutical composition comprises two ormore monoclonal antibodies or antigen-binding portions thereof asdescribed herein (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10 or more antibodies orantigen-binding portions thereof). In some embodiments, the compositioncomprises two or more monoclonal antibodies that specifically bind tothe same allergen. In some embodiments, the composition comprises two ormore monoclonal antibodies that specifically bind to different epitopesof the same allergen. In some embodiments, the composition comprises twoor more monoclonal antibodies that specifically bind to two or moredifferent allergens. It will also be appreciated by a person of ordinaryskill in the art that for a particular type or class of allergen, e.g.,a type of food allergen such as a peanut allergen, there can be morethan one substance (e.g., peptide or protein) within that type or classof allergen that induces an allergic response. In some embodiments, acomposition comprises two or more monoclonal antibodies thatspecifically bind to different allergens within a particular type orclass of allergen, e.g., two or more different peptides or proteins thatare allergens of the same type or class (e.g., two or more differentproteins that are peanut allergens). In some embodiments, thecomposition comprises two or more monoclonal antibodies thatspecifically bind to the same first allergen and further comprises oneor more monoclonal antibodies that specifically bind to a secondallergen.

Guidance for preparing formulations can be found in any number ofhandbooks for pharmaceutical preparation and formulation that are knownto those of skill in the art. See, e.g., Remington: The Science andPractice of Pharmacy, 21st Edition, Philadelphia, Pa. LippincottWilliams & Wilkins, 2005.

In some embodiments, the pharmaceutical composition further comprisesone or more pharmaceutically acceptable carriers, adjuvants, and/orvehicles appropriate for the particular route of administration forwhich the composition is to be employed. In some embodiments, thecarrier, adjuvant, and/or vehicle is suitable for intravenous,intramuscular, oral, intraperitoneal, transdermal, topical, orsubcutaneous administration. In some embodiments, the pharmaceuticalcomposition is formulated for intravenous or subcutaneousadministration. Methods of formulating antibodies for injection orinfusion (e.g., subcutaneous or intramuscular injection or byintravenous infusion) are also described in the art. See, e.g., US2013/0209465,

Pharmaceutically acceptable carriers are well-known in the art. See,e.g., Handbook of Pharmaceutical Excipients (5^(th) ed., Ed. Rowe etal., Pharmaceutical Press, Washington, D.C.). Examples ofpharmaceutically acceptable carriers include, but are not limited to,aqueous solutions, e.g., water or physiologically compatible bufferssuch as Hanks's solution, Ringer's solution, or physiological salinebuffer.

Typically, a pharmaceutical composition for use in in vivoadministration is sterile. Sterilization can be accomplished accordingto methods known in the art, e.g., heat sterilization, steamsterilization, sterile filtration, or irradiation.

Dosages and desired drug concentration of pharmaceutical compositions ofthe disclosure may vary depending on the particular use envisioned. Thedetermination of the appropriate dosage or route of administration iswell within the skill of one in the art. Suitable dosages are alsodescribed in Section V above.

In some embodiments, an antibody formulation comprising one or moreallergen-specific monoclonal antibodies or antigen-binding portionsthereof as disclosed herein are provided. In some embodiments, theantibody formulation comprises an antibody or antigen-binding portionthereof; and a buffer.

In some embodiments, the buffer is an acetate, citrate, histidine,succinate, phosphate, or hydroxymethylaminomethane buffer. In someembodiments, the antibody formulation further comprises one or moreadditional excipients such as a salt, a surfactant,polyol/disaccharide/polysaccharides, amino acids, and/or an antioxidant.In some embodiments, the antibody formulation comprises a surfactantsuch as polysorbate 80 (Tween 80), polysorbate 20 (Tween 20), orpoloxamer 188. In some embodiments, the antibody formulation comprises apolyol/disaccharide/polysaccharide such as mannitol, sorbitol, sucrose,trehalose, or dextran 40. In some embodiments, the antibody formulationcomprises a salt such as sodium chloride. In some embodiments, theantibody formulation comprises an amino acid such as glycine orarginine. In some embodiments, the antibody formulation comprises anantioxidant such as ascorbic acid, methionine, orethylenediaminetetraacetic acid (EDTA). In some embodiments, theantibody formulation is a lyophilized formulation. In some embodiments,the antibody formulation is a liquid formulation.

Kits

In some embodiments, kits comprising one or more allergen-specificmonoclonal antibodies or antigen-binding portions thereof as disclosedherein, or a pharmaceutical composition comprising one or moreallergen-specific monoclonal antibodies or antigen-binding portionsthereof as disclosed herein, are provided. In some embodiments, the kitcomprises a monoclonal antibody as described herein, e.g., as disclosedin Section IV above. In some embodiments, the kit comprises two or moremonoclonal antibodies or antigen-binding portions thereof (e.g., 2, 3,4, 5, 6, 7, 8, 9, 10 or more antibodies or antigen-binding portionsthereof) as described herein. In some embodiments, the kit is for use ina method of reducing one or more allergy symptoms in a subject (e.g.,allergy symptoms due to an allergy to a food allergen, a plant allergen,a fungal allergen, an animal allergen, a dust mite allergen, a drugallergen, a cosmetic allergen, or a latex allergen). In someembodiments, the kit is for use in a method of reducing one or moreallergy symptoms in a subject having a food allergy, e.g., a peanutallergy. In some embodiments, the kit is for use in a method of reducingone or more allergy symptoms in a subject having a fungal allergy. Insome embodiments, the kit is for use in a method of reducing one or moreallergy symptoms in a subject having an allergy to two or more allergens(e.g., two or more food allergens, e.g., a peanut allergen and a treenut allergen).

In some embodiments, the kits can further comprise instructionalmaterials containing directions (i.e., protocols) for the practice ofthe methods of this invention (e.g., instructions for using the kit fortreating an allergy). While the instructional materials typicallycomprise written or printed materials they are not limited to such. Anymedium capable of storing such instructions and communicating them to anend user is contemplated by this invention. Such media include, but arenot limited to electronic storage media (e.g., magnetic discs, tapes,cartridges, chips), optical media (e.g., CD ROM), and the like. Suchmedia may include addresses to internet sites that provide suchinstructional materials.

VII. Examples

The following examples are offered to illustrate, but not to limit, theclaimed invention.

Example 1—Single Cell Transcriptomic Analysis of Human IgE ProducingCells and Their Antibodies from Allergic Individuals Abstract

IgE antibodies provide immunity from helminth infections, but also cancause life-threatening allergic reactions. Despite their importance tohuman health, these antibodies and the cells that produce them remainenigmatic due to their scarcity in humans; much of our knowledge oftheir properties is derived from model organisms. Herein the isolationof IgE producing B cells from the blood of individuals with foodallergies is described, followed by a detailed study of their propertiesby single cell RNA sequencing (scRNAseq). It has been discovered thatIgE B cells are deficient in membrane immunoglobulin expression and thatthe IgE plasmablast state is more immature than that of other antibodyproducing cells. Through recombinant expression of monoclonal antibodiesderived from single cells, cross-reactive IgE antibodies specific forboth major peanut allergens Ara h 2 and Ara h 3 were discovered andcharacterized; these are among the highest affinity native humanantibodies discovered to date. Surprisingly, an example of convergentevolution in unrelated individuals who independently evolved nearlyidentical antibodies to peanut allergens was found. Finally, it wasdiscovered that splicing within B cells of all isotypes revealspolarized germline transcription of the IgE, but not IgG4, isotype aswell as several examples of biallelic expression of germlinetranscripts. These results offer insights into IgE B celltranscriptomics, clonality and regulation, provide a striking example ofadaptive immune convergence, and offer an approach for acceleratingmechanistic disease understanding by characterizing a rare B cellpopulation underlying IgE-mediated disease at single cell resolution.

Introduction

The IgE antibody class is the least abundant of all isotypes in humansand plays an important role in host defense against parasitic worminfections (1), but it can also become misdirected towards otherwiseharmless antigens. Food allergies are one example of this misdirection,where symptoms ranging from urticaria to potentially fatal anaphylaxisresult from the degranulation of mast cells and basophils induced by therecognition of allergic food proteins by surface-bound IgE antibodies.Despite this central role in immunity and allergic disease, human IgEantibodies remain poorly characterized due to their scarcity (2). Bulkepitope mapping experiments have revealed that IgE antibodies arepolyclonal and epitopes are heterogeneous (3); however, individuals withthe same allergy tend to recognize a core set of one or a few allergenicproteins (4). Recent studies applying bulk fluorescence activated cellsorting (FACS) immunophenotyping (5,6) and immune repertoire deepsequencing (7) have inferred IgE B cell origins, while studiesperforming peanut allergen specific single cell sorting (8,9) havedescribed clonal families to which IgE antibodies belong. However, nonehave successfully isolated single IgE producing cells or the pairedheavy and light chain sequences that comprise individual IgE antibodies,leaving unanswered questions as to the functional properties of suchantibodies, transcriptional programs of these cells, and the degree towhich any of these features are shared across individuals. Similarly,there is a lack of knowledge, but growing interest, surrounding the IgG4isotype due to its potential role in mediating the reduced clinicalallergen reactivity that accompanies immunotherapy and early allergenexposure through antigen blocking (10). Here we report the firstsuccessful isolation and transcriptomic characterization of single IgEand IgG4 producing B cells from humans. We combined single cell RNAsequencing (scRNA-seq) with functional antibody assays to elucidatemechanisms underlying the regulation of IgE and to discover highaffinity, cross-reactive peanut specific antibodies in unrelatedindividuals.

Characterization of Single B Cells from Peripheral Blood

We performed scRNA-seq on B cells isolated from the peripheral blood offood allergic individuals, which enabled us to characterize each cell'sgene expression, splice variants, and heavy and light chain antibodysequences (FIG. 6). Fresh peripheral blood from six peanut allergicindividuals was first separated into plasma and cellular fractions;plasma was stored and later used for allergen-specific IgE concentrationmeasurements (FIGS. 7A-7C), while the cellular fraction was enriched forB cells prior to FACS (see Materials and Methods). CD19+ B cells of allisotypes were sorted exclusively based on immunoglobulin surfaceexpression, but with an emphasis on maximizing IgE B cell capture (FIGS.8A-8C). Because cellular identity was determined from scRNA-seq ratherthan a complex, many-color gating scheme, we were able to sort andcapture cells with high sensitivity. This approach makes the prospect ofIgE B cell capture accessible for many laboratories without stringentrequirements on FACS gate purity.

Single cells were sorted into 96 well plates, processed using a modifiedversion of the SmartSeq2 protocol (11) and sequenced on an IlluminaNextSeq with 2×150 bp reads to an average depth of 1-2 million reads percell (FIGS. 9A-9G). Sequencing reads were independently aligned andassembled to produce a gene expression count table and reconstructantibody heavy and light chains, respectively (FIG. 6, Materials andMethods). Using STAR (12) for alignment also facilitated the assessmentof splicing within single cells. Cells were stringently filtered toremove those of low quality, putative basophils, and those lacking asingle productive heavy and light chain, yielding a total of 973 cellsfor further analysis (Materials and Methods). The isotype identity ofeach cell was determined by its productive heavy chain assembly, whichavoids misclassification of isotype based on FACS immunoglobulin surfacestaining (FIG. 8B), a problem which is especially pervasive for IgE Bcells due to CD23, the “low-affinity” IgE receptor that captures IgE onthe surface of non-IgE B cells (6).

Principal component analysis of normalized gene expression followingbatch effect correction (FIGS. 9A-9G and Materials and Methods)separated cells into two distinct clusters (FIG. 2A) identifiable asplasmablasts (PBs) and naive/memory B cells. PBs expressed the triad oftranscription factors BLIMP1 (PRDM1), XBP1, and IRF4 that drive plasmacell differentiation (13), as well as genes associated with antibodysecretion, such as the J chain, while naive and memory cells expressedthe canonical mature B cell surface marker CD20 (MS4A1), as well astranscription factor IRF8, which antagonizes the PB fate and insteadpromotes a germinal center response (14). Additional data corroboratedthis cell subtype assignment; PBs had greater FACS forward and sidescatter in agreement with their larger size and increased granularity,PB cDNA concentrations were higher following preamplification, and PBsexpressed more antibody heavy and light chain transcripts (FIGS.10A-10D).

We assessed isotype distribution within each B cell subtype and foundthat, in stark contrast to other isotypes, IgE B cells overwhelminglybelonged to the PB subtype (FIG. 2B-2C). This discovery is consistentwith observations of preferential differentiation of IgE B cells intoPBs in mice (15). Subtype proportions for other isotypes followedexpectations: IgM B cells, which are primarily naive, had the lowest PBpercentages, while IgA B cells had the highest in accordance with theirsecretory role in maintaining mucosal homeostasis. Interestingly, wefound that the number of circulating IgE B cells for each individualcorrelated with total plasma IgE levels (FIG. 7C); a similar phenomenonhas been noted in cases of hyper-IgE syndrome (16).

By clustering antibodies into clonal families (CFs) we were able toobserve elements of classical germinal center phenomenon such as somatichypermutation, class switching, and fate determination in our data.Antibody heavy chain sequences were first divided by V and J genes andwere clustered if their amino acid CDR3 sequences shared at least 75%similarity. Only 49 heavy chains formed CFs with multiple members,although this was not surprising given the vast diversity of potentialimmunoglobulin gene rearrangements (FIG. 2D). We found that in contrastto other isotypes, IgE and IgG4 were surprisingly clonal as over 20% ofIgE and IgG4 antibodies belonged to such multi-member CFs (FIG. 2E).Multi-member CFs were diverse; they contained between two and sixsequences, had variable isotype membership (node pattern), and had acomprehensive distribution of mutational frequency (node size). CFs werespecific to an individual (node shape), with the exception of one CF(CF1) that contained six heavily mutated IgE PB sequences: three eachfrom individuals PA12 and PA13, as discussed in depth later. Four CFsillustrated the two possible terminal differentiation pathways ofgerminal centers in that they contained both PBs and memory B cells.Other CFs contained cells belonging to multiple isotypes, with one ofparticular interest (CF3), discussed later, that contained an IgE PB andan IgG4 PB. Validating the premise of CFs as a collection of cells withsimilar origin, we found light chain CDR3 sequences were oftencomparable within families (FIG. 2G).

IgE antibodies varied widely in gene usage, CDR3 lengths, and mutationfrequency (FIGS. 3A-3C). There was moderate correlation between themutation frequency of heavy and light chains within single cells (FIG.3C), with evidence of selection via an enrichment of replacementmutations relative to silent mutations in the heavy chain CDR1 and CDR2that was absent in framework (FWR) regions. Light chains were similarlyenriched for replacement mutations in the CDR1 and, to a lesser degree,FWR1 (FIG. 3D). Compared to other isotypes, IgE B cells had a similardistribution of heavy chain mutation frequency, relative utilization ofthe lambda versus kappa light chains, and heavy chain V and J gene usage(FIGS. 11A-11K).

B Cell Intrinsic Factors Define IgE Cell State

To elucidate B cell intrinsic factors affecting PB activation, survival,and differentiation, we assessed genes differentially expressed betweenIgE PBs and PBs of other isotypes (FIG. 3E). A host of MHC genes wererobustly upregulated in IgE PBs, suggesting a more immaturetranscriptional program given the established loss of MHC-II during thematuration of PBs to plasma cells (17) (18) (19). FCER2 (CD23), the“low-affinity” IgE receptor was also highly upregulated, although itsprecise role within IgE PBs is unclear; autoinhibition of IgE productioncould result from membrane CD23-mediated co-ligation of membrane IgE(mIgE) and CD21 (20). IgE production could also be upregulated throughantagonistic effects of soluble CD23 (21), which is produced followingcleavage by ADAM10 (22), a disintegrin and metalloproteinasedomain-containing protein that we find is co-expressed in a subset ofIgE PBs. LAPTM5, a negative regulator of B cell activation, BCRexpression, and antibody production (23), was upregulated, whichsuggests compromised activation and proliferation capacity of IgE PBs.Downregulated genes included LILRB4 (24), galectin 1 (LGALS1), whichsupports plasma cell survival (25), and the 5100 proteins S100A4,S100A6, and S100A10, which may indicate reduced proliferative andsurvival signaling (26,27). One of the most significantly downregulatedgenes in IgE PBs was spleen associated tyrosine kinase (SYK), whichplays an essential role in BCR signal transduction (28) and is necessaryfor naïve differentiation into plasma cells and for memory B cellsurvival (29). Taken together, this gene expression program shows thatthe IgE PB cell state is immature relative to other PBs with weakenedactivation, proliferation, and survival capacity. It also provides apotential transcriptomic mechanism for the hypothesized short-lived IgEPB phenotype described in mouse models of allergy (15).

B cell intrinsic factors also regulate IgE production in murine modelsvia impaired memory formation (30,31). Indeed, we found human IgE Bcells belonging to the naïve/memory subset were deficient in heavy chainmembrane immunoglobulin exon splicing compared to other common isotypes.Furthermore, membrane exon splicing was detected at low levels innon-IgE PBs, but not in IgE PBs (FIGS. 3F and 3G). In fact, the absenceof mIgE splicing rendered us unable to assess the relative utilizationof the two splice variants of mIgE known to have distinct signalingcharacteristics (32,33). The lack of mature mIgE transcripts could beexplained by poor processing of pre-mRNA (34) and is consistent with lowIgE surface protein we measured by FACS; indeed, mIgE surface proteinlevels on true IgE B cells did not exceed those of some non-IgE B cellspresumably displaying surface IgE as a result of CD23-mediated capture(FIG. 8B). Together, these results suggest that the scarcity ofcirculating memory IgE B cells in vivo could result from impairedmembrane IgE expression that compromises IgE B cell entry into thememory compartment and/or memory B cell survival. Murine studies supportsuch a hypothesis, having shown IgE responses are reduced by removal ormodification of mIgE domains, but augmented by the exchange of thesedomains for those of IgG1 (35).

Characterization of Peanut-Specific IgE and IgG4 Antibodies

Surprisingly, our clonal analysis produced one CF of cells belonging tomultiple individuals (CF1, FIGS. 2F and 2G), which contained three IgEPBs from individual PA12 and three IgE PBs from individual PA13. Theantibodies produced by these six cells were highly similar as allutilized the IGHV3-30*18 and IGHJ6*02 heavy chain genes as well as theIGKV3-20*01 and IGKJ2*01 light chain genes, with pairwise CDR3 aminoacid sequence identity ranging from 65% to 94% for the heavy chain and70% to 100% for the light chain. These antibodies were also highlymutated and enriched in replacement mutations within the complementaritydetermining regions of both chains (FIG. 4A). In fact, compared to allother class switched antibodies, these were amongst the most mutated:the heavy chains were in the 76th percentile or above for mutationfrequency, while all of the light chains were in the 96th percentile orabove (FIG. 4B).

We recombinantly expressed the six IgE antibodies belonging to thisconvergent clonal family in order to assess whether they bind thenatural forms of the major allergenic peanut (Arachis hypogaea) proteinsAra h 1, Ara h 2, or Ara h 3. Of all characterized peanut allergens, Arah 2 is the most commonly recognized by allergic individuals and is themost clinically relevant both in terms of immunological response (36)and discriminating allergic status (37,38). Using an indirect ELISA as asemi-quantitative screen for binding, we found these six antibodiesbound strongly to Ara h 2, moderately to Ara h 3, and very weakly to Arah 1 (FIGS. 12A-12H). We then used biolayer interferometry to determinedissociation constants of each antibody for Ara h 2 and Ara h 3, withresulting affinities of 17 picomolar (pM) to sub-pM for Ara h 2 andmicromolar to sub-nanomolar for Ara h 3 (FIG. 4C and FIGS. 12A-12G).These affinities are comparable to some of the highest affinity nativehuman antibodies discovered for pathogens such as HIV, influenza, andmalaria (39-43). Additionally, high affinity to multiple peanutallergens should be advantageous if such antibodies or variants thereofwere to be used therapeutically as blocking antibodies intended tooutcompete endogenous IgE for allergenic protein, an approach recentlyshown to be efficacious for treatment of cat allergy (44).

To investigate the degree to which each chain and the mutations thereinaffect the binding properties, we recombinantly expressed eight variantsof antibody PA13P1H08, each with one or more regions in the heavy and/orlight chain reverted to the inferred naïve rearrangement. Reversion ofthe heavy chain CDR3 was performed based on the aforementioned heavychain V and J gene segments as well as the IGHD4-11*01 D gene andinferred nontemplated nucleotides TYCT between the V and D genes.Reversion of the light chain CDR3 was performed based on theaforementioned light chain V and J genes. Retaining the native heavychain while swapping the light chain with another kappa light chainabrogated binding to both allergenic proteins, while reverting bothchains eliminated Ara h 3 specificity and dramatically reduced Ara h 2affinity (FIG. 4C). Reverting only the heavy or light chain reduced theaffinity to Ara h 2 and Ara h 3, but disproportionately; light chainmutations contributed more to Ara h 3 affinity than did heavy chainmutations. We also found a synergistic contribution of heavy chainmutations to affinity as independent reversion of the CDR1, CDR2, CDR3or framework region(s) each caused minor decreases in affinity.Interestingly, reversion of the heavy chain CDR2 increased Ara h 3affinity, while only marginally decreasing Ara h 2 affinity. Theseresults indicate that the naively recombined antibody sequence iscapable of binding the most clinically relevant peanut allergen Ara h 2,but mutations, especially in the light chain, are necessary forgenerating high affinity antibodies which are cross-reactive towards Arah 3. More broadly, this shows that two unrelated individuals produced anidentically rearranged naive B cell that bound Ara h 2 and underwentclass switching, affinity maturation, differentiation, and clonalexpansion, eventually resulting in the presence of multiple circulatingIgE PBs secreting high affinity antibodies with cross-reactivity towardsAra h 3.

We also expressed antibodies from two other CFs. CF2 contained three IgEPBs from individual PA16 (two of which were identical), but theirrecombinantly expressed antibodies did not bind Ara h 1, 2, or 3, whichwas unsurprising given this individual had low plasma peanut-specificIgE levels as well as IgE specific to other allergens (FIGS. 7A-7C). Onthe other hand, CF3 contained an IgE PB (PA15P1D05) and IgG4 PB(PA15P1D12), the recombinantly expressed antibodies from which did notbind Ara h 1, but bound Ara h 3 with nanomolar affinity and Ara h 2 withsub-nanomolar affinity (FIGS. 12A-12H). Interestingly, these twoantibodies utilize the same light chain V gene and a highly similarheavy chain V gene (IGHV3-30-3*01) as the six convergent antibodies ofCF1, which provides additional support for the importance of these Vgenes in Ara h 2 binding. Moreover, the presence of peanut-specific IgEand IgG4 in the same CF within an allergic individual provides a uniqueexample of antagonist cell fate given the roles of IgE and IgG4 inallergic reactivity and potentially decreased sensitization,respectively.

Polarized Germline Transcription and Class Switch Priming

Tailored responses of the adaptive immune system are possible in partdue to the ability of activation-induced cytidine deaminase (AID) toinitiate class switch recombination (CSR) in B cells, leading to theproduction of antibodies with specific effector functions. CSR ispreceded by cytokine-induced germline transcription, where nonproductivegermline transcripts (GLTs) that contain an 1-exon, switch (S) region,and heavy chain constant region exons guide AID to the S region (45).Importantly, GLT processing is necessary for CSR (46,47) and canonicallyresults in two species: an intronic S region lariat and a maturepolyadenylated transcript consisting of the I exon spliced to theconstant region exons (48). In our scRNA-seq data, we observe multiplesplice isoforms of the latter, where the proximal constant region exonserves as the exclusive splice acceptor for multiple splice donors. IgEhad the largest number of distinct GLTs at five (FIG. 5A and FIG. 14),which we confirmed through Sanger sequencing; these were expressed innumerous cells of varying isotypes and across all individuals, but atnonuniform frequencies (FIG. 5A). The 1-exon was the most common splicedonor site (FIG. 5A, GLT #1) and it is known that 1-exons can providemultiple splice donors (49-51), but εGLT splice donors within the switchregion were also observed.

We found independent evidence for multiple IgE GLT splice donors in apreviously published scRNA-seq dataset from murine B cells harvested 24h after simulation to class switch (52) (FIG. 15). We also assessedvariation in the isotypes expressing εGLTs. The IgG4 isotype had thehighest proportion of cells expressing an εGLT (FIG. 5B), while IgE Bcells themselves also commonly expressed εGLTs. The remainder ofisotypes had fairly low expression of εGLTs.

GLT production is not limited to the IgE locus; we extended our analysisto all isotypes, enabling the construction of a global class switchpriming state diagram (FIG. 5D) that illustrates the fraction of cellsof each isotype that produce a GLT of their own (self) or anotherisotype. We observe that, in contrast to IgE, we find almost no IgG4 GLTexpression in these allergic individuals. We also observe elevated IgG2GLT production, which can be explained by splicing of the CH1 IgG2 exonto an upstream lincRNA. Interestingly, we observe that GLT expressionarising from the alternate allele is common, as evidenced by commonexpression of IgM GLTs as well as GLTs of other isotypes upstream of aclass switched isotype (signal below the diagonal in FIG. 5D). Mirroringthe landscape of human class switching (53), we observe the trend forGLT production to be higher for proximal downstream isotypes rather thandistant downstream isotypes. Unlike previous reports (54), we found thatcells with GLT expression tend to be polarized towards the expression ofa single GLT isotype, although we did not detect any non-self GLTproduction in most cells (FIG. 5E).

The study of B lymphocyte transcriptomes at single cell resolutionoffers other advantages; for example, we discovered multiple instancesof biallelic GLT expression though heavy chain constant region haplotypephasing in single B cells from in individuals who had heterozygoussingle nucleotide variants within these loci. An example of this processthat demonstrates biallelic εGLT expression is shown in FIG. 5C.

Characterization of Tree Nut-Specific IgE and IgG4 Antibodies

Given that some subjects had plasma IgE against other allergens inaddition to peanut (FIG. 7A), we assessed whether recombinant monoclonalantibodies from subjects PA11, PA12, PA13, PA14, PA15, and PA16 bound toallergen extracts derived not only from peanut, but other allergents aswell, including cashew, pistachio, latex, BSA, soy, sesame, milk, egg,almond, pine nuts, pecan, walnut, hazelnut, and macadamia. The resultsof an indirect ELISA are shown in FIG. 12H. A total of 89 antibodieswere tested, although only those with any OD value above 0.25 are shownin FIG. 12H. Antibodies not depicted include: PA12P1D04, PA12P1G02,PA16P1B09, PA16P1E11, PA16P1E12, PA11P1C01, PA11P1C12, PA11P1C06,PA11P1C08, PA11P1D07, PA11P1E08, PA11P1F10, PA11P1F02, PA11P1G06,PA11P1G07, PA13P2H10, PA15P1C03, PA15P1E01, PA15P1E02, PA13P1C01,PA13P1C09, PA13P1D02, PA13P1E06, PA14P1C10, PA14P1C12, PA14P1C02,PA14P1C04, PA14P1C06, PA14P1C07, PA14P1C08, PA14P1D10, PA14P1D02,PA14P1D07, PA14P1D09, PA14P1E10, PA14P1E11, PA14P1E12, PA14P1E04,PA14P1E06, PA14P1E08, PA14P1E09, PA14P1F10, PA14P1F11, PA14P1F05,PA14P1F07, PA14P1G01, PA14P1G11, PA14P1G12, PA14P1G03, PA14P1H01,PA14P1H11, PA14P1H12, PA14P1H02, PA14P1H05, PA14P1H09, PA12P3C05,PA12P3C09, PA12P3D11, PA12P3D09, PA12P3E06, PA12P3E07, PA12P3F02,PA12P3F07, PA13P3G04, PA14P3F10, PA14P3F02, PA14P3H10, PA14P3H12,PA12P4G03, and PA12P4G06.

As shown in FIG. 12H, several antibodies exhibited specificity formultiple tree nut allergens. The antibody PA14P3H08 bound strongly topecan, walnut, hazelnut, and macadamia allergens; the antibody PA11P1D11bound to pecan, walnut, and macadamia allergens; the antibodiesPA11P1E01, PA11P1C11, and PA11P1C03 each strongly bound to cashew andpistachio allergens; and the antibody PA11P1C04 more weakly bound tocashew and pistachio allergens. Some antibodies exhibited specificityfor both peanut and tree nut allergens. For example, the antibodyPA11P1G10 strongly bound to both pecan and walnut allergens and alsobound (albeit more weakly) to peanut allergen, while the antibodyPA12P4D02 strongly bound to peanut allergen and more weakly bound towalnut allergen while not binding natural peanut allergen Ara h 2. Otherantibodies exhibited specificity for a single tree nut allergen. Forexample, antibody PA11P1G04 bound to pistachio allergen, PA11P1F03 boundto pecan allergen, and PA11P1D12 bound to macadamia allergen.Furthermore, additional peanut-specific antibodies were discoveredduring these experiments. Antibodies PA12P3E09 and PA12P3E11 boundpeanut extract with little to no binding to natural peanut allergen Arah 2, while antibodies PA12P1D02, PA12P1G11, PA13P1H03, PA12P3C01, andPA12P3E04 bound strongly to both peanut extract and natural peanutallergen Ara h 2.

Conclusion

Using scRNA-seq, we provide the first transcriptomic characterization ofcirculating human IgE B cells and the antibodies they produce. Our datasuggests two mechanisms underlying IgE regulation in humans: membraneimmunoglobulin expression deficiency and an IgE PB gene expressionprogram suggestive of weakened activation, proliferation, and survivalcapacity. These results are largely consistent with extensive studies ofmIgE signaling and IgE memory in murine models of allergy, and provideevidence supporting the use of animal models for this disease. (55-59).Furthermore, the ability to capture GLT splice variant, polarization,and biallelic expression information within single B cells presents anexciting application of scRNA-seq for future mechanistic studies of GLTand CSR.

Insight into convergent evolution of high affinity antibodies inunrelated individuals can guide vaccine design and lead to strategiesfor population-level passive immunity; it is also a process that hasbeen argued to occur in response to a number of pathogens such asinfluenza (60), HIV (43), and Streptococcus pneumoniae (61). Here wefound a striking case of convergence where two unrelated individualsproduced high affinity, cross-reactive, peanut-specific antibodiescomprised of identical gene rearrangements within respective heavy andlight chains. A third individual has Ara h 2-specific antibodies thatutilize a similar heavy V gene and the same light chain V gene. Althoughthis is a small sample size, there is evidence supporting the importanceof these genes within the peanut-allergic population more broadly: oneindependent dataset of IgE heavy chain sequences from peanut allergicindividuals (62) contains IgE heavy chains that utilize identical V andJ genes and share at least 70% CDR3 identity with one or more of the sixconvergent antibodies in our dataset (FIG. 16); another dataset (9)contains Ara h 2 specific antibodies belonging to IgG and IgM B cellsthat utilize similar IGHV3-30 genes.

Cross-inhibition experiments with purified allergens and plasma IgE haveshown that cross-reactivity of IgE antibodies may also be common withinpeanut allergic individuals (63) and the antibodies we have isolatedhere offer a clear example of these findings. Furthermore, the fact thatthese high affinity antibodies were being produced by secretory IgE PBsfound in circulation contributes to an understanding of how minuteamounts of allergen are capable of eliciting severe allergic reactions.We also expect that either these antibodies themselves or engineeredvariants of them may find application as therapeutics; recent clinicalresults have shown that engineered allergen-specific IgG antibodies canbe administered to humans and provide effective treatment forcat-whisker allergies, perhaps by outcompeting the native IgE forantigen (44).

Methods Study Subjects

All study subjects were consented and screened through the Stanford IRBapproved-protocol. Participants were eligible if they had a peanutallergy confirmed by an oral food challenge and board certifiedallergist. Peanut allergic individuals with reported reactivity topeanut ranged in age from 8 to 17, and in some cases exhibitedsensitivities to other food allergens (FIGS. 7A-7C).

Plasma IgE Measurement and B Cell Isolation

Both plasma and cellular fractions were extracted from up to 45 mL offresh peripheral blood collected in K2 EDTA tubes. For plasmaextraction, blood was transferred to 15 mL falcon tubes and spun at 1600g for 10 min. The upper plasma layer was extracted, transferred to 2 mLEppendorf protein LoBind tubes and spun again at 16000 g to furtherpurify the plasma fraction. The resulting supernatant was moved to freshtubes before being put on dry ice and later transferred to −80° C.Allergen-specific plasma IgE measurements were performed byCLIA-licensed Johns Hopkins University Dermatology, Allergy, andClinical Immunology (DACI) Reference Laboratory using the ImmunoCAPsystem. To purify B cells remaining after plasma extraction, RosetteSephuman B cell enrichment cocktail (Stemcell Technologies), a negativeselection antibody cocktail, was added after the plasma fraction wasreplaced with PBS+2% fetal bovine serum (FBS). After a 20 minincubation, the blood was then diluted two-fold with PBS+2% FBS beforebeing transferred to Sepmate 50 mL tubes (Stemcell Technologies)containing 15 mL Ficoll-Plaque PLUS (GE Healthcare Life Sciences). Anenriched B cell population was achieved after a 10 min, 1200 g spin withthe brake on and transferred a fresh tube. Residual red blood cells werethen removed using ACK lysis buffer (ThermoFisher) and cells were washedwith stain buffer (BD Biosciences). Cells were stained on ice with thefollowing BioLegend antibodies according to the manufacturer'sinstructions: PE anti-human IgE clone MHE-18, Brilliant Violent 421anti-human CD19 clone HIB19, APC anti-human IgM clone MHM-88, and AlexaFluor 488 anti-human IgG clone M1310G05. Cells were washed twice moreprior to sorting.

Flow Cytometry and Single Cell Sorting

Single cell sorts were performed on a FACSAria II Special Order ResearchProduct (BD Biosciences) with a 5 laser configuration (355, 405, 488,561, and 640 nm excitation). Fluorophore compensation was performedprior to each sort using OneComp eBeads (ThermoFisher), although minimalcompensation was required due to the fluorophore panel and laserconfiguration. Equivalent laser power settings were used for each sort.Cells were sorted using “single cell” purity mode into chilled 96 wellplates (Biorad HSP9641) containing lysis buffer (11) and ERCC syntheticRNA spike-in mix (ThermoFisher). Plates were spun and put on dry iceimmediately before storage at −80° C.

cDNA Generation, Library Preparation, and Sequencing

A modified version of the SmartSeq2 protocol (64) was used as previouslydescribed (11). In total, 1165 cells were sequenced across 5 runs using2×150 bp Illumina High Output kits on an Illumina NextSeq.

Sequencing Read Alignment, Gene Expression, and Splicing

Sequencing reads were aligned to the genome in order to determine geneexpression, identify splice variants, and assess read coverage. Toproduce the gene expression counts table, reads were first aligned tothe GRCh38 human genome using STAR v2.5.3a (12) run in 2-pass mode. Genecounts were then determined using htseq-count (65) run inintersection-nonempty mode. The GTF annotation file supplied to bothSTAR and htseq-count was the Ensembl 90 release manually cleaned oferroneous Ig transcripts e.g. those annotated as either a V gene orconstant region but containing both V gene and constant region exons.During STAR genome generation an additional splice junction file wasprovided that included splicing between all combinations of heavy chainCH1 exons and IGHJ genes to improve read mapping across these junctions.Gene expression was normalized using log 2 counts per million afterremoving counts belonging to ERCCs. Cells with fewer than 950 expressedgenes were excluded prior to analysis, as were putative basophils,identified by high FACS IgE, absent or poor quality antibody assemblies,and expression of histidine decarboxylase (HDC) and Charcot-Leydencrystal protein/Galectin-10 (CLC). Batch effects mostly affecting thenaive/memory B cell subset were noted between sorts by clustering usingPCA on the 500 most variable genes; this gene set was enriched in genesknown to be affected by sample processing such as FOS, FOSB, JUN, JUNB,JUND, HSPA8 (66). PCA following the exclusion of genes differentiallyexpressed between sort batches (Mann-Whitney test, p-value <0.01 afterBonferroni correction) yielded well-mixed populations within both thenaive/memory and PB cell clusters not biased by sort batch, individual,or sequencing library (FIG. 9G). For differential expression analysisbetween IgE and non-IgE PBs, genes expressed in at least 10 PBs wereanalyzed by voom-limma (67) with sort batch and sequencing library weresupplied as technical covariates. Constant region genes, such as IGHEand IGHA1, were excluded given these are differentially expressed bydesign of the comparison being made.

Analysis of splicing, including GLT expression, relied upon splicejunctions called by STAR. Junctions were discarded if they containedfewer than three unique reads and GLT splice donors were only consideredif observed in at least three cells. Biallelic expression of GLTs wasdetermined based on heterozygous expression of variants discoveredwithin heavy chain constant regions using bcftools (68).

Antibody Heavy and Light Chain Assembly

In addition to alignment, sequencing reads were also independentlyassembled in order to reconstruct full length heavy and light chaintranscripts. BASIC (69) was used as the primary assembler given itsintended use for antibody reconstruction, while Bridger (70), a wholetranscriptome assembler, was used as an alternative when BASIC did notassemble a functional heavy and/or light chain. The heavy chain isotypeor light chain type (lambda or kappa) was determined using a BLAST (71)database of heavy and light chain constant regions constructed from IMGTsequences (72). Here it is important to note the necessity of isotypedetermination using heavy chain transcript presence rather than FACSimmunoglobulin surface staining: only 30% of B cells in the IgE B cellsort gate were in fact producing IgE transcripts (FIG. 8B). This likelyresults from the presence of surface-bound IgE captured by CD23 onnon-IgE B cells, and while acid-washing can remove IgE bound by CD23(73), we avoided this harsh treatment in order minimize transcriptomicperturbations to the cells. Immunoglobulin variable domain gene segmentassignment was performed using IgBLAST (74) v1.8.0 using a database ofhuman germline gene segments from IMGT. IgBLAST output was parsed withChange-0 and mutation frequency was called with SHazaM (75). Cellswithout a single productive heavy and single productive light chain,which were all members of the naive/memory cell cluster, were excluded,leaving a final total of 973 cells. The workflow engine Snakemake (76)was used to execute these analysis pipelines.

Recombinant Antibody Expression

Select antibodies were expressed recombinantly for specificity andaffinity assays. All heavy chains were expressed as human IgG1, whilelight chains were expressed as either lambda or kappa as appropriate.Heavy and light chain sequences were synthesized by Genscript aftercodon optimization and were transiently transfected in HEK293-6E cells.Antibodies were purified with RoboColumn Eshmuno® A columns (EMDMillipore) and were confirmed under reducing and non-reducing conditionsby SDS-PAGE and by western blots with goat anti-human IgG-HRP and goatanti-human kappa-HRP or goat anti-human lambda-HRP as appropriate.

Functional Antibody Characterization

ELISAs were performed one of two ways. For antibodies derived from CF1,CF2, or CF3, purified peanut allergens were used to semi-quantitativelyassess peanut allergen binding. Purified natural Ara h 1 (NA-AH1-1), Arah 2 (NA-AH2-1) and Ara h 3 (NA-AH3-1), purchased from IndoorBiotechnologies, were immobilized overnight at 4° C. using 50 μL at aconcentration of 2 ng/μL. Following 3 washes, wells were blocked with100 μL of PBST (ThermoFisher)+2% BSA for 2 hours. After two washes, 100μL of primary antibodies were incubated for 2 hours at a concentrationof 2 ng/μL in blocking buffer. Following 4 washes, 100 μL of rabbitanti-human HRP (abcam #ab6759) or rabbit anti-mouse HRP (abcam #ab6728)secondary antibodies were incubated for 2 hours at a dilution of 1/1000in blocking buffer. After 5 washes, 150 μL of 1-Step ABTS SubstrateSolution (ThermoFisher) was added to the wells. Color development wasmeasured at 405 nm on a plate reader after 8-20 min and reported ODvalues are after subtraction of signal from no-antibody wells. Negativecontrols included immobilized BSA as an antigen, as well as a humanisotype control primary antibody (abcam #ab206195). One random IgM/IgKantibody we recombinantly expressed (PA12P4H3) also did not exhibit anybinding. Positive controls consisted of monoclonal mouse antibodies2C12, 1C4, and 1E8 (Indoor Biotechnologies) specific for Ara h 1, Ara h2, and Ara h 3, respectively.

For ELISAs testing recombinant antibodies against a broad panel ofallergen extracts, the following was performed. First, the allergenswere obtained. Raw nut allergens, sesame seeds, peanuts, non-fat drymilk, and soy flour were purchased at a local grocery market, whilespray-dried whole egg was purchased from the National Institute ofStandards and Technology (RM 8445), and liquid latex containing naturalrubber centrifuged latex and water was obtained from Amazon. Ifnecessary, a mortar and pestle was used to grind solid allergens,following which 100 mg was added to a 2 mL Eppendorf Protein LoBind tubealong with a 5 mm stainless steel bead and 1.7 mL PBS. A TissueLysersystem (Qiagen) was used to homogenize the sample at 30 Hz for 10 min.Subsequently, the samples were spun for 20 min at 20000 g and 4° C. Theaqueous layer was then transferred to a fresh tube. The proteinconcentrations of these allergen extracts were then determined using thePierce 660 nm protein assay kit (ThermoFisher) in microplate formataccording to the manufacturer's instructions. ELISAs were performed in384 well format according to the following steps. First, 20 μL of 15allergens and BSA were incubated overnight at 4° C. at a concentrationof 2 ng/μL each. The plate was then washed 3 times with 62.5 μL of1×PBST per well per wash using an Integra VIAFLO. Wells were thenblocked for 2 hrs using 50 μL of a blocking buffer consisting of 1×PBSTand 2% BSA. Next, 20 μL of recombinant antibodies were incubated at aconcentration of 2 ng/μL in blocking buffer. Following 4 washes, 20 μLof rabbit anti-human HRP (abcam #ab6759) diluted 1/1000 in blockingbuffer was incubated for 2 hours. Following 5 washes, 40 μL of ABTS wasadded and 405 nm plate absorbance was measured using the BioTek Neo2.

Kinetic characterization of antibody interactions with natural purifiedallergenic peanut proteins was achieved using biolayer interferometry ona ForteBio Octet 96 using anti-human IgG Fc capture (AHC) biosensorswith 1×PBST as the assay buffer. The assay was run with the followingprotocol: up to 600s baseline, 120-150s antibody load, 120-300sbaseline, associations of up to 300s, and variable length dissociationsthat lasted up to 30 min for high affinity antibody-antigeninteractions. Biosensors were regenerated by cycling between buffer andpH 1.5 glycine following each experiment. Antibodies were loaded at aconcentration of 10-25 nM, while optimal peanut protein concentrationswere determined experimentally (FIGS. 12A-12H). Data were processedusing ForteBio software using a 1:1 binding model and global fit afterreference sensor (ligand, but no analyte) subtraction.

Example 2—Consensus Ara h 2 Binding Motif for Monoclonal AntibodyPA13P1H08 Methods

Linear epitope mapping of the recombinant IgG1 PA13P1H08 antibody wasperformed against Ara h 2 and Ara h 3 sequences linked and elongatedwith neutral GSGSGSG linkers at the N- and C-termini to avoid truncatedpeptides. The linked antigen sequences were translated into linear 15amino acid peptides with a peptide-peptide overlap of 14 amino acids.The resulting Ara h 2 and Ara h 3 peptide microarray contained 668different peptides printed in duplicate (1,336 peptide spots) as well as90 spots of influenza virus hemagglutinin (HA) peptide YPYDVPDYAGframing the microarray as internal quality controls.

The microarray was first subjected to 15 min pre-swelling in washingbuffer (PBS, pH 7.4 with 0.05% Tween 20), followed by 30 min in blockingbuffer (Rockland blocking buffer MB-070). The microarray was incubatedwith the PA13P1H08 IgG1 antibody at a concentration of 1 μg/ml inincubation buffer (washing buffer with 10% blocking buffer) for 16 h at4° C. with shaking at 140 rpm. The microarray was then stained withsecondary goat anti-human IgG (H+L) DyLight680 antibody (1:5000) andcontrol mouse monoclonal anti-HA (12CA5) DyLight800 antibody (1:2000)for 45 min in incubation buffer at room temperature. Read-out wasperformed with the LI-COR Odyssey Imaging System with the followingparameters: scanning offset 0.65 mm, resolution 21 μm, scanningintensities of 7/7 (red=700 nm/green=800 nm). Quantification of spotintensities and peptide annotation were done with PepSlide® Analyzer.

An identical copy of the peptide microarray was subjected to the aboveprocedure without incubation of the PA13P1H08 antibody. This served as acontrol to analyze background interactions of the secondary and controlantibodies with the 668 different peptides of both antigens.

Results

To assess whether the PA13P1H08 antibody, and by extension antibodiessimilar to PA13P1H08, could be binding linear peanut allergenepitope(s), we synthesized a microarray containing 15 amino acidpeptides from peanut allergens Ara h 2 and Ara h 3. We found secondaryand control antibody staining of the Ara h 2 and Ara h 3 peptidemicroarray did not highlight any background interactions that couldinterfere with the main assay (FIG. 17A). In contrast, we observed astrong PA13P1H08 antibody response with high signal-to-noise ratios tothe very similar consensus motifs DSYGRDPYSPS, YSPSQDPYSPS, andPDRRDPYSPS of Ara h 2. The similar responses were found in a region withmultiple repeat units with the common DPYSPS motif (FIGS. 17B and 17C).This motif occurs three times in the Ara h 2 isoform Ara h 2.0201 (SEQID NO:708; WHO/IUIS allergen nomenclature; Uniprot ID: Q6PSU2-1).

Example 3—Aspergillus-Specific Antibodies Derived from Human IgE B Cells

This example describes the generation and characterization ofAspergillus-specific antibodies derived from human IgE B cells. Themethods of Example 1 were used to obtain Aspergillus-specificantibodies, with the following differences: the blood from which B cellswere isolated originated from a subject (subject number 10033201) withallergic reactivity to Aspergillus, rather than a food allergy.Accordingly, the subject's plasma was tested for Aspergillus-specificIgE as well as for common food allergens and as shown in FIG. 18, plasmaIgE levels indicate Aspergillus sensitization but no confounding foodsensitizations.

Functional assays (ELISAs) were performed as described in Example 1 tosemi-quantitatively assess the obtained antibodies' specificity forstatically grown, defatted, powdered, and dried Aspergillus speciespurchased from Stallergenes Greer as well as recombinant Aspergillusfumigatus antigen Asp f 1 purchased from Indoor Biotechnologies. Asshown in FIG. 19, the monoclonal antibody clones 1003320101_D6,1003320105_D6, and 1003320107_F8 each exhibited specific binding toAspergillus fumigatus. Clone 1003320107_F3 exhibited specific binding topurified recombinant allergen Aspergillus fumigatus 1 (rAsp f 1).Furthermore, clone 1003320107_C5 specifically bound to each of theallergens Aspergillus fumigatus, Aspergillus niger, and Aspergillusnidulans and to rAsp F 1. These results demonstrate that the methoddisclosed herein for generating antibodies from single IgE- orIgG4-expressing human B cells is able to robustly isolateallergen-specific antibodies independent of the type of allergicdisease.

Example 4—Milk Allergen-Specific Antibodies Derived from Human IgE BCells

This example describes the generation and characterization ofmilk-specific antibodies derived from human IgE B cells. The methods ofExample 1 were used to obtain milk-specific antibodies, with thefollowing differences: the blood from which B cells were isolatedoriginated from a subject (PA01) with allergic reactivity to cow's milkbut not to other food allergens. The subject's plasma was tested forcommon food allergens, including milk. As shown in FIG. 18, plasma IgElevels from the subject indicate milk sensitization but no otherconfounding food sensitizations.

Functional assays (ELISAs) were performed as described in Example 1 tosemi-quantitatively assess the specificity of the obtained IgE and IgG4antibodies. As shown in FIG. 20, the monoclonal antibody clonesPA01P2D09, PA01P2D04, PA01P2B12, PA01P2B05, and PA01P2D10 each stronglybound to milk extract, while the antibody PA01P2E08 bound moderately.These results demonstrate that the method disclosed herein forgenerating antibodies from single IgE- or IgG4-expressing human B cellsis able to robustly isolate allergen-specific antibodies independent ofthe type of allergic disease.

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TABLE 1 Sequence Listing SEQ ID NO Sequence Description    1QVQLVNSGGGVVQPGRSLRLSCVASGFTFSTFGIHWVRQAPGKGLEWVAVHeavy chain variable region forISNDGEKSESADSVKGRFTPSRDNSKNTVYLQMNNLRVEDTAVYYCAKVLclone PA13P1H08 and N-R variant DYSRYSYYYGMDVWGQGTTVIVSS of PA13P1H08   2 GFTFSTFG CDR-H1 for clone PA13P1H08 andPA13P1H08 variants N-R, rCDR2-N, rCDR3-N, rFWRs-N    3 ISNDGEKSCDR-H2 for clone PA13P1H08 and for PA13P1H08 variants N-R,rCDR1-N, rCDR3-N    4 AKVLDYSRYSYYYGMDV CDR-H3 for clone PA13P1H08 andPA13P1H08 variants N-R, rCDR1-N, rCDR2-N    5EIVLTQSPGTLSLSPGGRGTLSCRTSQTINNAHLAWYQHKPGQAPRLLIYLight chain variable region forGSSERATGVPDRFSGSGSGSDFTLTISSLEAEDFAVYYCQHYGRSPPYTFclone PA13P1H08 and PA13P1H08 GPGTKLDIK variants R-N, rCDR1-N, rCDR2-N,rCDR3-N, rFWRs-N    6 QTINNAH CDR-L1 for clone PA13P1H08 andPA13P1H08 variants R-N, rCDR1-N, rCDR2-N, rCDR3-N, rFWRs-N    7 GSSCDR-L2 for clone PA13P1H08 and PA13P1H08 variants R-N, rCDR1-N,rCDR2-N, rCDR3-N, rFWRs-N    8 QHYGRSPPYT CDR-L3 for clone PA13P1H08 andPA13P1H08 variants R-N, rCDR1-N, rCDR2-N, rCDR3-N, rFWRs-N    9QVQLVDSGGGVVQPGKSLRLSCVGSGFTFRTFGIHWVRQAPGKGLEWVAVClone PA13P1E10 - Heavy chainISNDGGNSASADSVKGRFTTSRDNSKNTVYLQINSLRPEDTAIYYCAKVL variable regionDYSAFSYYYGMDVWGQGTTVIVSS   10 GFTFRTFG Clone PA13P1E10 - CDR-H1   11ISNDGGNS Clone PA13P1E10 - CDR-H2   12 AKVLDYSAFSYYYGMDVClone PA13P1E10 - CDR-H3   13EIVLTQSPGTLSLSPGERGTLSCRTSQPISRAHLAWYQHKAGQAPRLLIYClone PA13P1E10 - light chainGSTERAAGIPERFSGGGSGSDFTLTISSLEAEDFAVYYCQHYGRSPPYTF variable regionGQGTKVEIK   14 QPISRAH Clone PA13P1E10 - CDR-L1   15 GSTClone PA13P1E10 - CDR-L2    8 QHYGRSPPYT Clone PA13P1E10 - CDR-L3   16QVQLVESGGGVVQPGGSLTLSCVGSGFTFSHYAIHWVRQAPGKGLEWVAVClone PA12P3F10 - Heavy chainISNVGTTRDYADSLKGRLTISRENSQSTVFLQMNSLRADDTAIYYCAKVL variable regionDYSEFHYYYGLDVWGQGTAVAVSS   17 GFTFSHYA Clone PA12P3F10 - CDR-H1   18ISNVGTTR Clone PA12P3F10 - CDR-H2   19 AKVLDYSEFHYYYGLDVClone PA12P3F10 - CDR-H3   20EIVLTQSPGTLSLSPGQRVTLSCRVSQAIPTMYVAWYQQRPGQAPRLLIYClone PA12P3F10 - light chainGTSSRATGIPDRFSGGGSGTDFTLTINRLEPEDIAVYYCQHYSNSPPYTF variable regionGPGTKLEIK   21 QAIPTMY Clone PA12P3F10 - CDR-L1   22 GTSClone PA12P3F10 - CDR-L2   23 QHYSNSPPYT Clone PA12P3F10 - CDR-L3   24QEQLVESGGGVVHPGGSLRLSCVASAFTFNRFGMHWVRQAPGKGLEWVAVClone PA13P3G09 - Heavy chainISNDGRSQDYADSVKGRFIISRDNSKNTLYLQLNSLRFEDTAVYYCAKVL variable regionDYSIFYYYFGLDVWGQGTTVTVSS   25 AFTFNRFG Clone PA13P3G09 - CDR-H1   26ISNDGRSQ Clone PA13P3G09 - CDR-H2   27 AKVLDYSIFYYYFGLDVClone PA13P3G09 - CDR-H3   28EVVLTQSPGSLSLSPGERATLSCRAGQSLSSKFLAWYQHKPGQAPRLLIYClone PA13P3G09 - light chainGASTRATGVPDRFSGSGSGTDFSLIISRVEPEDFAVYYCQHYGDSPPYTF variable regionGQGTKVEMK   29 QSLSSKF Clone PA13P3G09 - CDR-L1   30 GASCDR-L2 for clone PA13P3G09 and PA13P1H08 variants R-R, N-R   31QHYGDSPPYT Clone PA13P3G09 - CDR-L3   32QVQLVESGGGVVQPGKSLRLSCAASAFTFRRFAMHWVRQAPGKGLEWVAVClone PA12P3D08 - Heavy chainISDNGLREDYEDSVKGRFTISRDNSQNTLYLQMNGLRAEDTAVYYCAKVL variable regionDYNEYSLYFGMDVWGQGTTVTVSS   33 AFTFRRFA Clone PA12P3D08 - CDR-H1   34ISDNGLRE Clone PA12P3D08 - CDR-H2   35 AKVLDYNEYSLYFGMDVClone PA12P3D08 - CDR-H3   36EVVLTQSPATLSLSPGERATLSCRTSQAISNNFLAWYQQRPGQPPRLLIYClone PA12P3D08 - light chainASSRRATDTPDRFTGSGSGTDFTLTITRLEPEDFAVYFCQYYSDSPPYTF variable regionGPGTKLEIK   37 QAISNNF Clone PA12P3D08 - CDR-L1   38 ASSClone PA12P3D08 - CDR-L2   39 QYYSDSPPYT Clone PA12P3D08 - CDR-L3   40QVQLEESGGGVVQPGKSLRLSCVASAFTFKRFAMHWVRQAPGKGLEWVAVClone PA12P1C07 - Heavy chainISDNGLREDYEDSVKGRFTISRDNSKDTLYLQMNSLRPEDTAIYYCAKVL variable regionDYSEYSLYFGMDVWGQGTTVLVSS   41 AFTFKRFA Clone PA12P1C07 - CDR-H1   34ISDNGLRE Clone PA12P1C07 - CDR-H2   42 AKVLDYSEYSLYFGMDVClone PA12P1C07 - CDR-H3   43EIVLTQSPAILSLSPGDRATLSCRTSQTVNSNFLAWYQQKPGQAPRLLIYClone PA12P1C07 - light chainGASRRAIDIPDRFTGSGSGTEFTLTIARLEPEDFAVYSCQHYSDSPPYTF variable regionGQGTKLEIK   44 QTVNSNF Clone PA12P1C07 - CDR-L1   30 GASClone PA12P1C07 - CDR-L2   45 QHYSDSPPYT Clone PA12P1C07 - CDR-L3   46QVHLVESGGGVVQPGRSLGLSCAASGFTFNYYAIHWVRQAPGKGLEWVAVClone PA15P1D12 - Heavy chainVSFDGNIIYYADSVKGRFNISRDNSKNTVNLQMNSLRADDTAVYYCVRDG variable regionEYCSGGNCYWGDFDYWGQGTLVTVSP   47 GFTFNYYA Clone PA15P1D12 - CDR-H1   48VSFDGNII Clone PA15P1D12 - CDR-H2   49 VRDGEYCSGGNCYWGDFDYClone PA15P1D12 - CDR-H3   50EIVLTQSPGTLSLSPGERATLSCRASQSISSEYLTWFQQKPGQAPRLLIYClone PA15P1D12 - light chainGAFNRATGIPDRFSGSGSGTDFTLTISSLEPEDFAVYYCQQYANWWTFGQ variable regionGTKVEIK   51 QSISSEY Clone PA15P1D12 - CDR-L1   52 GAFClone PA15P1D12 - CDR-L2   53 QQYANWWT Clone PA15P1D12 - CDR-L3   54QVHLVESGGGVVQPGRSLGLSCVASGFTFNYYAIHWVRQAPGKGLEWVAVClone PA15P1D05 - Heavy chainVSFDGNIIYYADSVKGRFNISRDNSKNTVNLQMNSLRPDDTAVYYCVRDG variable regionEYCSGGNCYWGDFDHWGQGSLVTVSP   47 GFTFNYYA Clone PA15P1D05 - CDR-H1   48VSFDGNII Clone PA15P1D05 - CDR-H2   55 VRDGEYCSGGNCYWGDFDHClone PA15P1D05 - CDR-H3   56EIVLTQSPATLSLSPGERATLSCRASQSISSEYLTWFQQKPGQAPRLLIYClone PA15P1D05 - light chainGAFNRATGIPDRFSGSGSGTDFTLTISSLEPEDFAVYYCQQYANWWTFGQ variable regionGTKVEIK   51 QSISSEY Clone PA15P1D05 - CDR-L1   52 GAFClone PA15P1D05 - CDR-L2   53 QQYANWWT Clone PA15P1D05 - CDR-L3   57QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVHeavy chain variable region ISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKVLfor R-R and R-N variants  DYSNYYYYYGMDVWGQGTTVTVSS of PA13P1H08   58GFTFSSYG CDR-H1 for PA13P1H08 variants R- R, R-N, rCDR1-N   59 ISYDGSNKCDR-H2 for PA13P1H08 variants R- R, R-N, rCDR2-N   60 AKVLDYSNYYYYYGMDVCDR-H3 for PA13P1H08 variants R- R, R-N, rCDR3-N   61EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYLight chain variable for GASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPPYTFregion R-R and N-R variants  GQGTKLEIK of PA13P1H08   62 QSVSSSYCDR-L1 for PA13P1H08 variants R- R, N-R   63 QQYGSSPPYTCDR-L3 for PA13P1H08 variants R- R, N-R   64QVQLVNSGGGVVQPGRSLRLSCVASGFTFSSYGIHWVRQAPGKGLEWVAVHeavy chain variable region forISNDGEKSESADSVKGRFTPSRDNSKNTVYLQMNNLRVEDTAVYYCAKVLPA13P1H08 variant rCDR1-N DYSRYSYYYGMDVWGQGTTVIVSS   65QVQLVNSGGGVVQPGRSLRLSCVASGFTFSTFGIHWVRQAPGKGLEWVAVHeavy chain variable region forISYDGSNKESADSVKGRFTPSRDNSKNTVYLQMNNLRVEDTAVYYCAKVLPA13P1H08 variant rCDR2-N DYSRYSYYYGMDVWGQGTTVIVSS   66QVQLVNSGGGVVQPGRSLRLSCVASGFTFSTFGIHWVRQAPGKGLEWVAVHeavy chain variable region forISNDGEKSESADSVKGRFTPSRDNSKNTVYLQMNNLRVEDTAVYYCAKVLPA13P1H08 variant rCDR3-N DYSNYYYYYGMDVWGQGTTVIVSS   67QVQLVESGGGVVQPGRSLRLSCAASGFTFSTFGMHWVRQAPGKGLEWVAVHeavy chain variable region forISNDGEKSYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKVLPA13P1H08 variant rFWRs-N DYSRYSYYYGMDVWGQGTTVTVSS   68GCCTCCACACAGAGCCCATCCGTCTTCCCCTTGACCCGCTGCTGCAAAAANucleotide sequence for IgE heavyCATTCCCTCCAATGCCACCTCCGTGACTCTGGGCTGCCTGGCCACGGGCT chain constant regionACTTCCCGGAGCCGGTGATGGTGACCTGGGACACAGGCTCCCTCAACGGGACAACTATGACCTTACCAGCCACCACCCTCACGCTCTCTGGTCACTATGCCACCATCAGCTTGCTGACCGTCTCGGGTGCGTGGGCCAAGCAGATGTTCACCTGCCGTGTGGCACACACTCCATCGTCCACAGACTGGGTCGACAACAAAACCTTCAGCGTCTGCTCCAGGGACTTCACCCCGCCCACCGTGAAGATCTTACAGTCGTCCTGCGACGGCGGCGGGCACTTCCCCCCGACCATCCAGCTCCTGTGCCTCGTCTCTGGGTACACCCCAGGGACTATCAACATCACCTGGCTGGAGGACGGGCAGGTCATGGACGTGGACTTGTCCACCGCCTCTACCACGCAGGAGGGTGAGCTGGCCTCCACACAAAGCGAGCTCACCCTCAGCCAGAAGCACTGGCTGTCAGACCGCACCTACACCTGCCAGGTCACCTATCAAGGTCACACCTTTGAGGACAGCACCAAGAAGTGTGCAGATTCCAACCCGAGAGGGGTGAGCGCCTACCTAAGCCGGCCCAGCCCGTTCGACCTGTTCATCCGCAAGTCGCCCACGATCACCTGTCTGGTGGTGGACCTGGCACCCAGCAAGGGGACCGTGAACCTGACCTGGTCCCGGGCCAGTGGGAAGCCTGTGAACCACTCCACCAGAAAGGAGGAGAAGCAGCGCAATGGCACGTTAACCGTCACGTCCACCCTGCCGGTGGGCACCCGAGACTGGATCGAGGGGGAGACCTACCAGTGCAGGGTGACCCACCCCCACCTGCCCAGGGCCCTCATGCGGTCCACGACCAAGACCAGCGGCCCGCGTGCTGCCCCGGAAGTCTATGCGTTTGCGACGCCGGAGTGGCCGGGGAGCCGGGACAAGCGCACCCTCGCCTGCCTGATCCAGAACTTCATGCCTGAGGACATCTCGGTGCAGTGGCTGCACAACGAGGTGCAGCTCCCGGACGCCCGGCACAGCACGACGCAGCCCCGCAAGACCAAGGGCTCCGGCTTCTTCGTCTTCAGCCGCCTGGAGGTGACCAGGGCCGAATGGGAGCAGAAAGATGAGTTCATCTGCCGTGCAGTCCATGAGGCAGCGAGCCCCTCACAGACCGTCCAGCGAGCGGTGTCTGTAAATCCCGGTAAA   69ASTQSPSVFPLTRCCKNIPSNATSVTLGCLATGYFPEPVMVTWDTGSLNGAmino acid sequence for IgE heavyTTMTLPATTLTLSGHYATISLLTVSGAWAKQMFTCRVAHTPSSTDWVDNK chain constant regionTFSVCSRDFTPPTVKILQSSCDGGGHFPPTIQLLCLVSGYTPGTINITWLEDGQVMDVDLSTASTTQEGELASTQSELTLSQKHWLSDRTYTCQVTYQGHTFEDSTKKCADSNPRGVSAYLSRPSPFDLFIRKSPTITCLVVDLAPSKGTVNLTWSRASGKPVNHSTRKEEKQRNGTLTVTSTLPVGTRDWIEGETYQCRVTHPHLPRALMRSTTKTSGPRAAPEVYAFATPEWPGSRDKRTLACLIQNFMPEDISVQWLHNEVQLPDARHSTTQPRKTKGSGFFVFSRLEVTRAEWEQKDEFICRAVHEAASPSQTVQRAVSVNPGK   70GCTTCCACCAAGGGCCCATCCGTCTTCCCCCTGGCGCCCTGCTCCAGGAGNucleotide sequence for IgG4CACCTCCGAGAGCACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCheavy chain constant regionCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACGAAGACCTACACCTGCAACGTAGATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGTCCAAATATGGTCCCCCATGCCCATCATGCCCAGCACCTGAGTTCCTGGGGGGACCATCAGTCTTCCTGTTCCCCCCAAAACCCAAGGACACTCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTGGTGGACGTGAGCCAGGAAGACCCCGAGGTCCAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTTCAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGGCCTCCCGTCCTCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCATCCCAGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAGGCTAACCGTGGACAAGAGCAGGTGGCAGGAGGGGAATGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACACAGAAGAGCCTCTCCCTGTCTCTGGGTAAA   71ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVAmino acid sequence for IgG4HTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESheavy chain constant regionKYGPPCPSCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKFGYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQHEGNVFSCSVMHEALNHYTQKSLSLSLGK   72QVQLVESGGGVVQPGRSLRLSCAASGFTFDTYGMHWVRQAPGKGPEWVAVClone PA14P1E12 - Heavy chainIWYDGTREDYADSVKGRFTVSRDNSKSTLFLQMNSLRADDTAVYYCAKEH variable regionNTYFSDHIGRVGGMDVWGQGTTVIVSS   73 GFTFDTYG Clone PA14P1E12 - CDR-H1   74IWYDGTRE Clone PA14P1E12 - CDR-H2   75 AKEHNTYFSDHIGRVGGMDVClone PA14P1E12 - CDR-H3   76DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQClone PA14P1E12 - light chainLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQVLQTP variable regionPWTFGQGTQVEIK   77 QSLLHSNGYNY Clone PA14P1E12 - CDR-L1   78 LGSClone PA14P1E12 - CDR-L2   79 MQVLQTPPWT Clone PA14P1E12 - CDR-L3   80QVQLVESGGGVVQPGRSLRLSCAGSGFTFNAYGLHWVRQAPGKGLEWVAGClone PA14P1E10 - Heavy chainIYYDGSNKYYADSVKGRFAISRDNSQNTLYLEMNSLRVEDTAVYYCAKAG variable regionPIASIGTRHTFDHWGQGTLVTVSS   81 GFTFNAYG Clone PA14P1E10 - CDR-H1   82IYYDGSNK Clone PA14P1E10 - CDR-H2   83 AKAGPIASIGTRHTFDHClone PA14P1E10 - CDR-H3   84DIVMTQSPDSLAVSLGERATINCKSSQSLLLNSNNKNYLAWYQQKPGQPPClone PA14P1E10 - light chainKLLIYWASTRESGVPGRFSGNGSVTDFTLTISGLQAEDVAVYYCHQYYTT variable regionSYTFGQGTKLEIK   85 QSLLLNSNNKNY Clone PA14P1E10 - CDR-L1   86 WASClone PA14P1E10 - CDR-L2   87 HQYYTTSYT Clone PA14P1E10 - CDR-L3   88QVQLVQSGAEVKKPGASVKISCKAVGYTFTSYYLHWVRQAPGQGLEWVGIClone PA14P1E11 - Heavy chainIDPSRGHRNYAQGFQGRVTMTSDTSTSTVYMDLGSLRSEDTAVYYCARAP variable regionARDHFDNWGQGTPVTVSP   89 GYTFTSYY Clone PA14P1E11 - CDR-H1   90 IDPSRGHRClone PA14P1E11 - CDR-H2   91 ARAPARDHFDN Clone PA14P1E11 - CDR-H3   92DIQMTQSPSSLAASVGDRVTINCQASQDIRNCLNWYQQQPGKAPKLLIYDClone PA14P1E11 - light chainASILETGVPSRFSGSGSGTDFTFSISSLQPEDIATYYCQQCEDLPLTFGP variable regionGSKVDIK   93 QDIRNC Clone PA14P1E11 - CDR-L1   94 DASClone PA14P1E11 - CDR-L2   95 QQCEDLPLT Clone PA14P1E11 - CDR-L3   96QVQLVQSGAEVKQPGSSVKVSCKASGGTFRNSALSWVRQAPGQGLEWMGGClone PA13P1C01 - Heavy chainIIPIFDTTNYAQEFQGRVTITADKSTTTAYMELSSLKSEDTAVYYCARGE variable regionGLPWLTYHYYGMDVWGQGTTVTVSS   97 GGTFRNSA Clone PA13P1C01 - CDR-H1   98IIPIFDTT Clone PA13P1C01 - CDR-H2   99 ARGEGLPWLTYHYYGMDVClone PA13P1C01 - CDR-H3  100QAVLTQPSSLSASPGASASLTCTLRSGINIGTDRIYWFQQKPGSPPQYLLClone PA13P1C01 - light chainTYKSDSDEQRGSGVPSRFSGSKDVSANAGILLISGLQSEDEADYYCMIWH variable regionSSAWVFGGGTKLTVL  101 SGINIGTDR Clone PA13P1C01 - CDR-L1  102 YKSDSDEClone PA13P1C01 - CDR-L2  103 MIWHSSAWV Clone PA13P1C01 - CDR-L3  104QVQLQQWGAGLLKPSETLSLICAVYGGSLSGYHWSWIRQPPGKGLQWIGEClone PA14P1F11 - Heavy chainISHSGNAKYNPSLKSRVSISVHMSKNEFYLNLTSVTAADTAVYYCARGYC variable regionSGGSCYYKFWGQGTLVTVSS  105 GGSLSGYH Clone PA14P1F11 - CDR-H1  106 ISHSGNAClone PA14P1F11 - CDR-H2  107 ARGYCSGGSCYYKF Clone PA14P1F11 - CDR-H3 108 QSVLTQPPSVSAAPGQKVTISCSGNSSNIGNNYVSWFQQLPGTAPKLLIYClone PA14P1F11 - light chainDNNKRPSGIPDRFSGSKSGTSATLGITGLQTGDEADYFCGTWDSSLRTGV variable regionFGGGTKLTVL  109 SSNIGNNY Clone PA14P1F11 - CDR-L1  110 DNNClone PA14P1F11 - CDR-L2  111 GTWDSSLRTGV Clone PA14P1F11 - CDR-L3  112EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSLClone PA14P1F10 - Heavy chainIYSGGSRTSYPDSVKGRFTISRDNSNSTLFLQMNSLRVEDTAVYYCAKGG variable regionSSWLKMDYWGQGTLVIVSS  113 GFTFSSYA Clone PA14P1F10 - CDR-H1  114 IYSGGSRTClone PA14P1F10 - CDR-H2  115 AKGGSSWLKMDY Clone PA14P1F10 - CDR-H3  116QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVQWYQHLPGTAPKLLIClone PA14P1F10 - light chainFANTNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSSLSGS variable regionGVFGGTKLTVL  117 SSNIGAGYD Clone PA14P1F10 - CDR-L1  118 ANTClone PA14P1F10 - CDR-L2  119 QSYDSSLSGSV Clone PA14P1F10 - CDR-L3  120QVQLVQSGAEVKKPGASVRVSCSSSGYTFTGYYIHWVRQAPGQGLEYMGRClone PA13P1C09 - Heavy chainINPHSGGTNYAQKFQGRVTMTRDTSTSTVYMELSSLRSDDTAVYYCAKEG variable regionTTAHIFNWFDPWGQGTLVTVSS  121 GYTFTGYY Clone PA13P1C09 - CDR-H1  122INPHSGGT Clone PA13P1C09 - CDR-H2  123 AKEGTTAHIFNWFDPClone PA13P1C09 - CDR-H3  124DIQMTQSPSSLSASVGDRVTITCRASQSINSYLNWYQQKPGKAPNLLIYTClone PA13P1C09 - light chainASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYTSLFTFGQ variable regionGTKLEIK  125 QSINSY Clone PA13P1C09 - CDR-L1  126 TASClone PA13P1C09 - CDR-L2  127 QQSYTSLFT Clone PA13P1C09 - CDR-L3  128QLQLQESGPGLVKPSETLSLTCTVSGVSINSTSYYWGWMRQPPGKGLEWIClone PA13P1D02 - Heavy chainGNIYYTGTTYYNPSLNRRVSISGDTSKNQFSLSLTSVTAADTAVYYCAGP variable regionVRRTVFGILLMESFDVWSQGTMVTVSS  129 GVSINSTSYY Clone PA13P1D02 - CDR-H1 130 IYYTGTT Clone PA13P1D02 - CDR-H2  131 AGPRRVTVFGILLMESFDVClone PA13P1D02 - CDR-H3  132AIQMTQSPSSLSASVGDRVTITCRASQAIRDDLGWFQQKPGKAPKLLIYTClone PA13P1D02 - light chainASTLQSGVPSRFSGGGSGTEFILTISSLQPEDIGTYYCLQDYGYPWTFGQ variable regionGTKVEIK  133 QAIRDD Clone PA13P1D02 - CDR-L1  126 TASClone PA13P1D02 - CDR-L2  134 LQDYGYPWT Clone PA13P1D02 - CDR-L3  135EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYALSWVRQAPGKGLEWVSAClone PA14P1E04 - Heavy chainISGRDASTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCTLFD variable regionYDSSGYFDFDYWGQGTLVTVSS  136 GFTFSNYA Clone PA14P1E04 - CDR-H1  137ISGRDAST Clone PA14P1E04 - CDR-H2  138 TLFDYDSSGYFDFDYClone PA14P1E04 - CDR-H3  139QSALTQPRSVSGSPGQSVTISCTGTGSDVGGYNYVSWYQHHPGKAPKLIIClone PA14P1E04 - light chainFDVTKRPSGVPDRFSGSKSGYTASLTISGLQAEDEAVYYCCSYANSYTGV variable regionFGTGTKVTVL  140 GSDVGGYNY Clone PA14P1E04 - CDR-L1  141 DVTClone PA14P1E04 - CDR-L2  142 CSYANSYTGV Clone PA14P1E04 - CDR-L3  143QVQLVESGGGVVQPGGSLRLSCAASGFTFSSHVMHWVRQAPGKGLEWVALClone PA14P1E06 - Heavy chainISLDGDDKYYADSVNGRVAISRDNSKNTLYLQVNSLRSDDTCVYYCARGG variable regionRWDYALDVWGQGTTVTVSS  144 GFTFSSHV Clone PA14P1E06 - CDR-H1  145 ISLDGDDKClone PA14P1E06 - CDR-H2  146 ARGGRWDYALDV Clone PA14P1E06 - CDR-H3  147EIVMTQSPATLSVSPGERATLSCRVSQSISNSLAWYQQKPGQVPRLLIYAClone PA14P1E06 - light chainASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQYNNWPRALTF variable regionGGGTKVEIK  148 QSISNS Clone PA14P1E06 - CDR-L1  149 AASClone PA14P1E06 - CDR-L2  150 QQYNNWPRALT Clone PA14P1E06 - CDR-L3  151QVQLVESGGGVVQPGRSLRLSCAASGFTFNDYAMHWVRQAPGKGPEWVAVClone PA11P1G06 - Heavy chainISYDGTNEYYMGSVKGRFTISRDNSKNMVNLQMNSLRPEDTAVYYCARDL variable regionAAWSRELLVFDQWGQGTLVTVSS  152 GFTFNDYA Clone PA11P1G06 - CDR-H1  153ISYDGTNE Clone PA11P1G06 - CDR-H2  154 ARDLAAWSRELLVFDQClone PA11P1G06 - CDR-H3  155ENVLTQSPGTLSLSPGEGATLSCRASQSVPNTYLAWYQQKPGQAPRLLIYClone PA11P1G06 - light chainGASSRAAGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGRSPGTFG variable regionQGTKVEIK  156 QSVPNTY Clone PA11P1G06 - CDR-L1   30 GASClone PA11P1G06 - CDR-L2  157 QQYGRSPGT Clone PA11P1G06 - CDR-L3  158QAQVVESGGGVVQPGTSLRLSCEPSGFTLSDYGIHWVRQPPGKGLEWVAVClone PA11P1G07 - Heavy chainIWHDGDRINYADSVKGRFTISRDESDKKVHLQMESLRTEDTAVYYCARGT variable regionLPRNCRGMRCYGEFDHYYYLDVWGTGTTVTVSS  159 GFTLSDYG Clone PA11P1G07 - CDR-H1 160 IWHDGDRI Clone PA11P1G07 - CDR-H2  161 ARGTLPRNCRGMRCYGEFDHYYYLDVClone PA11P1G07 - CDR-H3  162QSVLTQPPSVSGAPGQRVTISCTGHSSNIGANSDVHWYQQLPLRAPKLLIClone PA11P1G07 - light chainFGTINRASGVPDRFSGSRSGTSASLVISGLQPDDEADYYCQSYDRGLSAY variable regionVFGSGTRVDVL  163 SSNIGANSD Clone PA11P1G07 - CDR-L1  164 GTIClone PA11P1G07 - CDR-L2  165 QSYDRGLSAYV Clone PA11P1G07 - CDR-L3  166EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYWMSWVRQAPGKGLEWVANClone PA11P1G04 - Heavy chainIKQDGSEKDYVDSVKGRFTISRDNAKNSLYLQLNSLRAEDTAVYYCARER variable regionSTQSSSWYVSSYYSYYGMDVWGQGTTVTVSS  167 GFTFSSYW Clone PA11P1G04 - CDR-H1 168 IKQDGSEK Clone PA11P1G04 - CDR-H2  169 ARERSTQSSSWYVSSYYSYYGMDVClone PA11P1G04 - CDR-H3  170SSELTQDPAVSVALGQTVTITCQGDSLRSFYASWYQQKPGQAPVFVIYGKClone PA11P1G04 - light chainYNRPSGIPDRFSGSSSGNTASLTITGAQAEDEADYYCNSRDSSDNHLGVF variable regionGGGTKLTVL  171 SLRSFY Clone PA11P1G04 - CDR-L1  172 GKYClone PA11P1G04 - CDR-L2  173 NSRDSSDNHLGV Clone PA11P1G04 - CDR-L3  174QVQLVQSGSELRKPGASVKLSCRTSGYTFIHFAMNWLRQAPGQGLEWLGWClone PA11P1C11 - Heavy chainINTHSGNPTYAQGFTGRFVFSLDVSAGTAYLEISGLKAEDTAVYYCARER variable regionYFDFWGQGALVAVSS  175 GYTFIHFA Clone PA11P1C11 - CDR-H1  176 INTHSGNPClone PA11P1C11 - CDR-H2  177 ARERYFDF Clone PA11P1C11 - CDR-H3  178QSVLTQPPSASGTPGQRVTISCSGTSSNIGKNFLYWYQQVPGTAPKLLIYClone PA11P1C11 - light chainSSNQRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLSGWV variable regionFGGGTKVTVL  179 SSNIGKNF Clone PA11P1C11 - CDR-L1  180 SSNClone PA11P1C11 - CDR-L2  181 AAWDDSLSGWV Clone PA11P1C11 - CDR-L3  182EVQLVESGGDLVQPGGSLRLSCAASGFTFSNYWMNWVRQPPGKGLVWVSRClone PA11P1C12 - Heavy chainISGDGTGTSYADSVRGRFTISRDNAKSTLYLQVNSLSAEDTAVYYCTRDG variable regionGRDHPTPDAFDIWGQGTMVTVSS  183 GFTFSNYW Clone PA11P1C12 - CDR-H1  184ISGDGTGT Clone PA11P1C12 - CDR-H2  185 TRDGGRDHPTPDAFDIClone PA11P1C12 - CDR-H3  186DVVMAQSPLSLPVTLGQPASISCRSSQSLVHSDGNTYLNWFQQRPGQSPRClone PA11P1C12 - light chainRLIYKISNRDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQGTHWP variable regionRTFGQGTKLEIK  187 QSLVHSDGNTY Clone PA11P1C12 - CDR-L1  188 KISClone PA11P1C12 - CDR-L2  189 MQGTHWPRT Clone PA11P1C12 - CDR-L3  190QVQLQQWGAGLLKPSETLSLTCVVSGGSFSTHYWNWIRQSPGKGLEWIGEClone PA15P1G05 - Heavy chainINHSGNTNYNPSLTGRATISVATSKTQFSLRLNSVTAADTAVYFCARGPR variable regionLRYTAGRPLFDTWGQGTLVTVSS  191 GGSFSTHY Clone PA15P1G05 - CDR-H1  192INHSGNT Clone PA15P1G05 - CDR-H2  193 ARGPRLRYTAGRPLFDTClone PA15P1G05 - CDR-H3  194DIQMTQSPSTLSASVGDRVTITCRASQSISAFLAWYQQKPGKAPNLVIYKClone PA15P1G05 - light chainASSLDSGVPSTFSGSGSGTEYTLTISSLQPDDFATYYCQQYFSSPPTFGQ variable regionGTKVEMK  195 QSISAF Clone PA15P1G05 - CDR-L1  196 KASClone PA15P1G05 - CDR-L2  197 QQYFSSPPT Clone PA15P1G05 - CDR-L3  198QVQLQESGPGLVKPSQTLSLTCAVSGGSISSGGYYWSWIRQLPGKGLEWIClone PA14P1F05 - Heavy chainGYIYYSGSTSYNPSLKSRVTISVDTSKNQLSLNLSSVTAADTAVYNCARG variable regionRRISISGVVTPLFDYWGQGTLVTVSS  199 GGSISSGGYY Clone PA14P1F05 - CDR-H1  200IYYSGST Clone PA14P1F05 - CDR-H2  201 ARGRRISISGVVTPLFDYClone PA14P1F05 - CDR-H3  202DIQMTQSPSSVSASVGDRVTITCRASQGISSWLAWYQLKPGKAPKLLIYAClone PA14P1F05 - light chainASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANSVPLTFGG variable regionGTKVEIK  203 QGISSW Clone PA14P1F05 - CDR-L1  149 AASClone PA14P1F05 - CDR-L2  204 QQANSVPLT Clone PA14P1F05 - CDR-L3  205EVQLVESGGGLVKPGGSLRLSCAASGFTFSHYYLNWVRQAPGKGLEWVACClone PA14P1F07 - Heavy chainISDRSENVYYADSVKGRFTISRDNAKNSLFLQMNNLRAEDTAIYYCARDM variable regionRELRPSADYWGQGTLVTVSS  206 GFTFSHYY Clone PA14P1F07 - CDR-H1  207ISDRSENV Clone PA14P1F07 - CDR-H2  208 ARDMRELRPSADYClone PA14P1F07 - CDR-H3  209EIVLTQSPGTLSLSPGDRATLSCRASQSVDGNSLAWYQQKPGQAPRLLISClone PA14P1F07 - light chainGASTRATGIPDRFSGSGSGTDFTLTISRLEPEDFVLYHCQLYTVSPRYTF variable regionGQGTKLEIK  210 QSVDGNS Clone PA14P1F07 - CDR-L1   30 GASClone PA14P1F07 - CDR-L2  211 QLYTVSPRYT Clone PA14P1F07 - CDR-L3  212QLQLQESGPGLVKPSETLSLTCTVSGGSISSDNYYWGWIRQPPGKGPLWIClone PA14P3H12 - Heavy chainGTIFYNGDTYYNPSLKSQLNISVDPSKNQFSLKLTSVTAADTAIYYCTRH variable regionDSYSRGWYVTHWGQGTLVTVSS  213 GGSISSDNYY Clone PA14P3H12 - CDR-H1  214IFYNGDT Clone PA14P3H12 - CDR-H2  215 TRHDSYSRGWYVTHClone PA14P3H12 - CDR-H3  216EIVLTQSPATLSLFPGERATLSCRASQSVTSYLAWYQQKPGQAPRLLIYDClone PA14P3H12 - light chainASKRATGIPARFSGSGSGTDFTLTISSLEPEDFATYYCQQRSARQLFGGG variable regionTKVEIK  217 QSVTSY Clone PA14P3H12 - CDR-L1   94 DASClone PA14P3H12 - CDR-L2  218 QQRSARQL Clone PA14P3H12 - CDR-L3  219EVQLVESGGGLVKPGGSLRLSCVASGLTFRNAWMTWVRQAPGKGLEWVGRClone PA14P3H10 - Heavy chainIKSNVNGGTTDYAAPVRGRFTISRDDSRDTLYLQMNSLETEDTAMYYCTK variable regionDPPYTGGGYCQHWGLGTLVTVSS  220 GLTFRNAW Clone PA14P3H10 - CDR-H1  221IKSNVNGGTT Clone PA14P3H10 - CDR-H2  222 TKDPPYTGGGYCQHClone PA14P3H10 - CDR-H3  223EIVLTQSPATLSLSPGESATLSCRASQSVSSCLAWYQQKPGQAPRLLIYDClone PA14P3H10 - light chainSASTRAPGIPGRFSGGSGTDFTLAISSLEPEDFAVYYCQQCSNWPLTFGR variable regionGTRLEIK  224 QSVSSC Clone PA14P3H10 - CDR-L1   94 DASClone PA14P3H10 - CDR-L2  225 QQCSNWPLT Clone PA14P3H10 - CDR-L3  226QVQLQESGPGLVKPSQTLSLTCTVSGGSINTGAYYWSWIRQHPGKGLEWIClone PA11P1G10 - Heavy chainGYIYYSGSTYYNPSLKSRVTISKDTSKNQFSLRLTSVTAADTAVYYCVRE variable regionKLTGAPDNWGQGTLVAVSS  227 GGSINTGAYY Clone PA11P1G10 - CDR-H1  200IYYSGST Clone PA11P1G10 - CDR-H2  228 VREKLTGAPDNClone PA11P1G10 - CDR-H3  229DIQMTQSPSSLSASVGDRVTITCRASQGVSNYLAWFHQKPGKAPKSLIYAClone PA11P1G10 - light chainASTLHDGVPSSFSGSGSGTEFTLTISDLQPEHFGTYYCEQYNSYPFTFGP variable regionGTTVDFK  230 QGVSNY Clone PA11P1G10 - CDR-L1  149 AASClone PA11P1G10 - CDR-L2  231 EQYNSYPFT Clone PA11P1G10 - CDR-L3  232QVQLVQSGAEVKKPGSSVKVSCKASGGPLSSYNFIWVRQAPGQGLEWMGGClone PA13P2H10 - Heavy chainILPVFDTTNYAQKFQGRVTITADKATSTSYMELSSLTSEDTAVYYCARAV variable regionGGTHYYYYGLDVWGQGTTVAVSS  233 GGPLSSYN Clone PA13P2H10 - CDR-H1  234ILPVFDTT Clone PA13P2H10 - CDR-H2  235 ARAVGGTHYYYYGLDVClone PA13P2H10 - CDR-H3  236EIVMTQSPLSLPVTPGEPASISCRSSQSLLHGNGYNYVDWYLQRPGQPPQClone PA13P2H10 - light chainLLIYLGSRRASGVPDRFSGSGSGTDFTLKISRVEADDLGVYYCMQALQTR variable regionVTFGPGTKVDIK  237 QSLLHGNGYNY Clone PA13P2H10 - CDR-L1   78 LGSClone PA13P2H10 - CDR-L2  238 MQALQTRVT Clone PA13P2H10 - CDR-L3  239EVQLVQSGAEVKKPGESLKISCKGSGYSFMSYWIGWVRQKPGKGLEWMGIClone PA14P1H02 - Heavy chainIFPGDSDTRYSPSFQGHVTISADKSITTAYLQWNSLEASDTAIYYCATLD variable regionGDYWGRGTLVTVSS  240 GYSFMSYW Clone PA14P1H02 - CDR-H1  241 IFPGDSDTClone PA14P1H02 - CDR-H2  242 ATLDGDY Clone PA14P1H02 - CDR-H3  243DIVMTQSPDSLAVSLGERATINCRSSQSVLSSSSNKNYLGWYQQKPGQPPClone PA14P1H02 - light chainKLLIHWASTRAAGVPDRFSGSGTGTDFTLNISSLQAEDVAVYYCQQYHTT variable regionLPTFGQGTKLEIK  244 QSVLSSSSNKNY Clone PA14P1H02 - CDR-L1   86 WASClone PA14P1H02 - CDR-L2  245 QQYHTTLPT Clone PA14P1H02 - CDR-L3  246EVQLLESGGGLVQPGGSLRLSCAASGFTFRDSAMTWVRQAPGKGLEWVSTClone PA14P1H01 - Heavy chainISGNGDTTYYADSVKGRFSIFRDNSRNTLYVQMNSLRAEDTAVYYCARYG variable regionDHKGWFDSWGQGTLVTVSS  247 GFTFRDSA Clone PA14P1H01 - CDR-H1  248 ISGNGDTTClone PA14P1H01 - CDR-H2  249 ARYGDHKGWFDS Clone PA14P1H01 - CDR-H3  250ELVMTQSPASLSVSPGEGATVSCRASQSVGSNLAWYQQKPGQGPRLLIYGClone PA14P1H01 - light chainASTRATGVPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQYNNWPRTFGQ variable regionGTKVEIK  251 QSVGSN Clone PA14P1H01 - CDR-L1   30 GASClone PA14P1H01 - CDR-L2  252 QQYNNWPRT Clone PA14P1H01 - CDR-L3  253QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIClone PA14P1H09 - Heavy chainINPSDGSKTYAQKFQGRVTLTRDTSTSTVYMELSSLRSEDTAVYYCARGN variable regionGYSSSWYVNDYWGQGTLVTVSS   89 GYTFTSYY Clone PA14P1H09 - CDR-H1  254INPSDGSK Clone PA14P1H09 - CDR-H2  255 ARGNGYSSSWYVNDYClone PA14P1H09 - CDR-H3  256DIVLTQSPGTLSLSPGERATLSCRASQSLTNSNFAWYQQIPGQAPRLLIYClone PA14P1H09 - light chainGASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFVVYYCQQYGRSPITFG variable regionQGTRLEIK  257 QSLTNSN Clone PA14P1H09 - CDR-L1   30 GASClone PA14P1H09 - CDR-L2  258 QQYGRSPIT Clone PA14P1H09 - CDR-L3  259QVQLVESGGGVVQPGRSLRLSCAASGFTFTKYGMHWVRQAPGKGLEWVALClone PA12P3D11 - Heavy chainISYDGNNKYYADSVRGRVTISRDNSKNTLYLQMDSLRAEDTAVYYCARGQ variable regionDYPFWSGSTFEYWGQGTLVTVSS  260 GFTFTKYG Clone PA12P3D11 - CDR-H1  261ISYDGNNK Clone PA12P3D11 - CDR-H2  262 ARGQDYPFWSGSTFEYClone PA12P3D11 - CDR-H3  263QAVVTQEPSLTVSPGGTVTLTCGSTTGAVTGGHFPYWIQQKPGQAPRTLIClone PA12P3D11 - light chainYDATNRHSWTPARFSGSLLGGKAALTLSGAQPEDEADYYCLLSYSSATFL variable regionIFGGGTKLTVL  264 TGAVTGGHF Clone PA12P3D11 - CDR-L1  265 DATClone PA12P3D11 - CDR-L2  266 LLSYSSATFLI Clone PA12P3D11 - CDR-L3  267QVQLQESGPGLVKPSETLSLTCTVSGDSLSSGSYFWSWIRQPPGKGLEWIClone PA12P3F02 - Heavy chainGYISFRGDTNYNPSLKSRVIISLDKSKNQFSLRLSSMTPADTAVYYCARS variable regionPWIQSWSYYFDYWGQGTLVTVSS  268 GDSLSSGSYF Clone PA12P3F02 - CDR-H1  269ISFRGDT Clone PA12P3F02 - CDR-H2  270 ARSPWIQSWSYYFDYClone PA12P3F02 - CDR-H3  271DIQMTQSPSTVSASVGDRVTITCRASQRISSWLAWYQQKPGKAPKLLIYKClone PA12P3F02 - light chainASSLEGGVPSRFSGSGSGTEFTLTISSLQPDDFAIYYCQQYNGYPWTFGQ variable regionGTKVEIK  272 QRISSW Clone PA12P3F02 - CDR-L1  196 KASClone PA12P3F02 - CDR-L2  273 QQYNGYPWT Clone PA12P3F02 - CDR-L3  274QVQLQESGPGLVKPSGTLSLTCAVSGGSISTDNWWSWVRQPPNKGLEWIGClone PA12P3F07 - Heavy chainAIFQSGSTIYNPSLMSRVTISLDRSNNRFSLQLISVTAADTALYYCARAS variable regionFHYGSGNYFEYLGQGTLVTVSS  275 GGSISTDNW Clone PA12P3F07 - CDR-H1  276IFQSGST Clone PA12P3F07 - CDR-H2  277 ARASFHYGSGNYFEYClone PA12P3F07 - CDR-H3  278QSVLTQPPSVSAAPGQKVTISCSGSSSNVGTHHVSWYQQLPGTAPKLLIYClone PA12P3F07 - light chainENDKRPSGIPNRFSGSKSGTSATLAIIGLQTGDEADYYCGSWDSSLSAFW variable regionVFGGGTKLTVL  279 SSNVGTHH Clone PA12P3F07 - CDR-L1  280 ENDClone PA12P3F07 - CDR-L2  281 GSWDSSLSAFWV Clone PA12P3F07 - CDR-L3  282QVQLVQSGAEVKKPGASVKVACKASGYTFTRYAMHWVRQAPGQRLEWMGWClone PA14P1G03 - Heavy chainINAGNGNTKDSQKFQGRVTITRDTSASTVYMELSSLRSEDTAVYYCARGV variable regionPWGLGSYNFDYWGQGTLVSISS  283 GYTFTRYA Clone PA14P1G03 - CDR-H1  284INAGNGNT Clone PA14P1G03 - CDR-H2  285 ARGVPWGLGSYNFDYClone PA14P1G03 - CDR-H3  286QTVVTQEPSLTVSPGGTVTLSCASNTGAVTSGYYPYWFQQKPGQAPRTLIClone PA14P1G03 - light chainYETSNKHPWTPARFSGSLLGGKAALTLSGVQPEDEAEYCCLLYYGGTWVF variable regionGGGTKLTVL  287 TGAVTSGYY Clone PA14P1G03 - CDR-L1  288 ETSClone PA14P1G03 - CDR-L2  289 LLYYGGTWV Clone PA14P1G03 - CDR-L3  290EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYALSWVRQAPGKGLEWVSAClone PA14P1G01 - Heavy chainISGRDGNTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCTLYD variable regionYDSSGYFDFDYWGQGTLVTVSS  113 GFTFSSYA Clone PA14P1G01 - CDR-H1  291ISGRDGNT Clone PA14P1G01 - CDR-H2  292 TLYDYDSSGYFDFDYClone PA14P1G01 - CDR-H3  293QSALTQPRSVSGSPGQSVTISCTGTSSDVGGFNYVSWYQQHPGKAPKLMIClone PA14P1G01 - light chainFDVTQRPSGVPDRFSGSKSGNTASLTISGLQAEDEADYHCCSYANYYTGV variable regionFGTGTRVTVL  294 SSDVGGFNY Clone PA14P1G01 - CDR-L1  141 DVTClone PA14P1G01 - CDR-L2  295 CSYANYYTGV Clone PA14P1G01 - CDR-L3  296EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYWMHWVRQAPGKGLVWVSRClone PA16P1F09 - Heavy chainINSDGSNIRFADSVKGRFTFSRDNANNTLYLQMNSLRAEDTAVYYCARAS variable regionRTVYGDSPLSYGIDVWGQGTTVTVSS  183 GFTFSNYW Clone PA16P1F09 - CDR-H1  297INSDGSNI Clone PA16P1F09 - CDR-H2  298 ARASRTVYGDSPLSYGIDVClone PA16P1F09 - CDR-H3  299SYALTQPPSVSVSPGQTASITCSGDKLGNKFACWYQQKPGRSPVLVIYQDClone PA16P1F09 - light chainSQRPTGIPERFSGSNSGNTATLTISGTQAMDEADYYCQAWDSNTHVLFGG variable regionGTKLTVL  300 KLGNKF Clone PA16P1F09 - CDR-L1  301 QDSClone PA16P1F09 - CDR-L2  302 QAWDSNTHVL Clone PA16P1F09 - CDR-L3  303QVQLVESGGGVVQPGGSLRLSCAASGFTFSGYGMHWVRQAPGKGLEWVAFClone PA14P1G12 - Heavy chainFSFDGSNTDYVDSVKGRFTISGDNSKNTLYLQMNSLRAEDTAVYYCVRDI variable regionLVLPAAVSVFSGYYYGMDVWGQGTTVIVSS  304 GFTFSGYG Clone PA14P1G12 - CDR-H1 305 FSFDGSNT Clone PA14P1G12 - CDR-H2  306 VRDILVLPAAVSVFSGYYYGMDVClone PA14P1G12 - CDR-H3  307EIVLTQSPATLSLSPGERATLSCRASQSVRSYLAWYQQKPGQAPRLLIYDClone PA14P1G12 - light chainASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQHRSNWPITFGQ variable regionGTRLEIK  308 QSVRSY Clone PA14P1G12 - CDR-L1   94 DASClone PA14P1G12 - CDR-L2  309 QHRSNWPIT Clone PA14P1G12 - CDR-L3  310QVQLRVSGPGLVNPSETLSLTCIVSGDSLRDYYWSWIRQSPGKGLEWIGYClone PA11P1D12 - Heavy chainVTESGGAHYNPSLESRVTISVDASKTQFSLNLKSVTAADTAVYYCARDAY variable regionSSTWYTVGWFDPWGPGSLVTVSS  311 GDSLRDYY Clone PA11P1D12 - CDR-H1  312VTESGGA Clone PA11P1D12 - CDR-H2  313 ARDAYSSTWYTVGWFDPClone PA11P1D12 - CDR-H3  314EIVLTQSPATLSLSPGERATLSCRASQDVGVYLAWYQQKPGQAPRLIIYDClone PA11P1D12 - light chainASDRVSGVPARFTGSGSGTDFTLTITSLEPEDFAVYFCQQRTSGLTFGGG variable regionTTLEIK  315 QDVGVY Clone PA11P1D12 - CDR-L1   94 DASClone PA11P1D12 - CDR-L2  316 QQRTSGLT Clone PA11P1D12 - CDR-L3  317QVELVESGGGVVQPGRSLRLSCVASGFTFSDYGMHWVRQAPGKGLEWVAVClone PA11P1D11 - Heavy chainIWFDGSSKYYADSVKGRFTISRDDSKNTVFMQMNNVRVEDTAVYYCAREQ variable regionWLGTEYFQNWGQGTLVTVSS  318 GFTFSDYG Clone PA11P1D11 - CDR-H1  319IWFDGSSK Clone PA11P1D11 - CDR-H2  320 AREQWLGTEYFQNClone PA11P1D11 - CDR-H3  321EIVMTQSPATLSLFPGERATLSCRASQSVAGNLAWYQQKPGQAPRLLIYEClone PA11P1D11 - light chainASTRATDIPARFSGSGSETEFTLTISSLQSEDFAVYYCQQYKKWLITFGQ variable regionGTRLEIK  322 QSVAGN Clone PA11P1D11 - CDR-L1  323 EASClone PA11P1D11 - CDR-L2  324 QQYKKWLIT Clone PA11P1D11 - CDR-L3  325QLQLQQWGAGLVKPSETLSLTCTVSGGSLSGHFWSWIRQSPEKGLEWIGEClone PA12P1D02 - Heavy chainINHSGRKNYNPSLMIRVDISIDTSKNQFSMRMTSLTAADSAVYYCARVGR variable regionNIVDTDDAFDVWGRGTLVTVSS  326 GGSLSGHF Clone PA12P1D02 - CDR-H1  327INHSGRK Clone PA12P1D02 - CDR-H2  328 ARVGRNIVDTDDAFDVClone PA12P1D02 - CDR-H3  329EIVLTQSPGTLSLSPGDTVTLSCRASQTIDSIYLAWYQQRPGQAPRLLIYClone PA12P1D02 - light chainGASTRATGTPDRFSGGGSGTDFTLTITRLEPEDFAVYFCQQYGTSPPITF variable regionGRGTRLEIK  330 QTIDSIY Clone PA12P1D02 - CDR-L1   30 GASClone PA12P1D02 - CDR-L2  331 QQYGTSPPIT Clone PA12P1D02 - CDR-L3  332QVQLLQSGAEVKKPGASVKVSCKASGYTFTSYNIHWVRQAPGQSFEWMGWClone PA12P1D04 - Heavy chainIHVGNGETKYSQNFQDRVAITRDTSANTVYMELSPLRSEDTALYYCVRDH variable regionVTAIVVGLFDPWGQGTLVTVSS  333 GYTFTSYN Clone PA12P1D04 - CDR-H1  334IHVGNGET Clone PA12P1D04 - CDR-H2  335 VRDHVTAIVVGLFDPClone PA12P1D04 - CDR-H3  336QSALTQPASVSGSPGQSITISCSGTSTDVGAYKYVSWYQHHPGRSPKVILClone PA12P1D04 - light chainYEVDNRPSGVSIRFSGSKSGNTASLTISGLRAEDEADYYCSSFTSSSTWV variable regionFGGGTKVTVL  337 STDVGAYKY Clone PA12P1D04 - CDR-L1  338 EVDClone PA12P1D04 - CDR-L2  339 SSFTSSSTWV Clone PA12P1D04 - CDR-L3  340QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYSISWVRQAPGQGLEWMGGClone PA12P3E09 - Heavy chainIIPIFGSGSYAQKFQGRVTITADKSTSTAYMELSSLSSDDTAVYYCARGE variable regionSPSNFVYYGMDVWGQGTTVTVSS  341 GGTFSSYS Clone PA12P3E09 - CDR-H1  342IIPIFGSG Clone PA12P3E09 - CDR-H2  343 ARGESPSNFVYYGMDVClone PA12P3E09 - CDR-H3  344DIVLTQSPLSLPVTPGEPASISCRSSHSLLHSNGYNHLDWYLQKPGQSPQClone PA12P3E09 - light chainLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQALLVT variable regionFGPGTKVDIK  345 HSLLHSNGYNH Clone PA12P3E09 - CDR-L1   78 LGSClone PA12P3E09 - CDR-L2  346 MQALLVT Clone PA12P3E09 - CDR-L3  347EVKVVESGGGLVQPGGSLRLSCAASEFTFTYYWMSWIRQAPGKGLEWVANClone PA12P3E04 - Heavy chainVNGDATEKYYVDSVKGRFTISRDNPKKTVYLQMNSLRVEDTAVYYCARVG variable regionTTVVNDGFDLWGLGTMVTVSS  348 EFTFTYYW Clone PA12P3E04 - CDR-H1  349VNGDATEK Clone PA12P3E04 - CDR-H2  350 ARVGTTVVNDGFDLClone PA12P3E04 - CDR-H3  351SYVLTQSHSVSVAPGQTARITCGGENIGGKGVHWYQQKPGQAPLLVVSSDClone PA12P3E04 - light chainTGRRSVTPDRFSGSNSGDTATLIISRVEAGDEADYYCQVWDPTSEYVFGS variable regionGTKVTVL  352 NIGGKG Clone PA12P3E04 - CDR-L1  353 SDTClone PA12P3E04 - CDR-L2  354 QVWDPTSEYV Clone PA12P3E04 - CDR-L3  355QLQLQESGSGLVKPSQTLSLTCAVSGGSISSGDYSWSWIRQPPGKGLEWIClone PA12P3E07 - Heavy chainGFRYYSGTTFYNPSLESRLTISIDRSTNQFSLQLTSVTAADTAVYFCASF variable regionRPLLRFLDPEGLFEYWGQGILVTVSS  356 GGSISSGDYS Clone PA12P3E07 - CDR-H1  357RYYSGTT Clone PA12P3E07 - CDR-H2  358 ASFRPLLRFLDPEGLFEYClone PA12P3E07 - CDR-H3  359ELVMTQSPATLSVSPGARATLSCRASPGANSHLAWYQQKPGQAPRLLIYGClone PA12P3E07 - light chainASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQYNDWPYTFGQ variable regionGTKLEIK  360 PGANSH Clone PA12P3E07 - CDR-L1   30 GASClone PA12P3E07 - CDR-L2  361 QQYNDWPYT Clone PA12P3E07 - CDR-L3  362QVQLVQSGAAVKKPGASVRISCEASGYTFTGYNIHWVRQAPGQGLEWMGWClone PA12P3E06 - Heavy chainVNPNNGGTKFAQKFEGWVTMTVATSINTVYMELTGLKSGDTAVYFCARDH variable regionGDSFDQWGQGTLVTVSS  363 GYTFTGYN Clone PA12P3E06 - CDR-H1  364 VNPNNGGTClone PA12P3E06 - CDR-H2  365 ARDHGDSFDQ Clone PA12P3E06 - CDR-H3  366EIVLTQSPDTLSLSPGDRATLSCRASHSLNNDYLAWYQHRPGQAPRLLIYClone PA12P3E06 - light chainGTSHGATGIPDRFSGSGSGTDFTLTISRLETEDFAVYYCHHYGKSLFPFG variable regionPGTKVDIK  367 HSLNNDY Clone PA12P3E06 - CDR-L1   22 GTSClone PA12P3E06 - CDR-L2  368 HHYGKSLFP Clone PA12P3E06 - CDR-L3  369EVQLVESGGGLVQPGGSLKLSCAASGFTFSGSAMHWVRQASGKGLEWVGRClone PA14P1E08 - Heavy chainIRSKANTYATAYAASVKGRFTISRDDSKNTAYLQMNSLKTEDTAVYYCTR variable regionKHTSGWYDRGGDVWGQGTTVTVSS  370 GFTFSGSA Clone PA14P1E08 - CDR-H1  371IRSKANTYAT Clone PA14P1E08 - CDR-H2  372 TRKHTSGWYDRGGDVClone PA14P1E08 - CDR-H3  373DIQMTQSPSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYDClone PA14P1E08 - light chainASNLETGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQYDNLPLTFGG variable regionGTKVEIK  374 QDISNY Clone PA14P1E08 - CDR-L1   94 DASClone PA14P1E08 - CDR-L2  375 QQYDNLPLT Clone PA14P1E08 - CDR-L3  376EVQLLESGGGLVQPGGSLRLSCAASGFTVSNYAMSWVRQAPGKGLEWVSAClone PA14P1E09 - Heavy chainISGSGGSTYYADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCAIDC variable regionTVTDAPLSYWGQGTLVTVSS  377 GFTVSNYA Clone PA14P1E09 - CDR-H1  378ISGSGGST Clone PA14P1E09 - CDR-H2  379 AIDCTVTDAPLSYClone PA14P1E09 - CDR-H3  380AIQMTQSPSSLSPSVGDRVTITCRASQGIRNDLGWYQQKPGKAPKLLIYAClone PA14P1E09 - light chainASSLQSGVPSRFSGGGSGTDFTLTISSLQPEDFATYYCLQDYNYPRTFGQ variable regionGTKVEIK  381 QGIRND Clone PA14P1E09 - CDR-L1  149 AASClone PA14P1E09 - CDR-L2  382 LQDYNYPRT Clone PA14P1E09 - CDR-L3  383QVQLEQSGAEVRKPGSSVKVSCKASGTTFSNHAMSWVRQAPGQGLEWMGGClone PA14P1H05 - Heavy chainIIPLVDKSMYALKFQGRVTITADESRNTVYMELSSLGSEDTAVYYCARSF variable regionADITTFGFVVNFHYYYTLDVWGQGTPVTVSS  384 GTTFSNHA Clone PA14P1H05 - CDR-H1 385 IIPLVDKS Clone PA14P1H05 - CDR-H2  386 ARSFADITTFGFVVNFHYYYTLDVClone PA14P1H05 - CDR-H3  387NFMLTQPHSVSESPGKTVTISCTRSSGSIADNYVQWFQQRPGSAPTTLIYClone PA14P1H05 - light chainEDNRRPSGVPDRFSGSVDSSSNSASLTISGLKPEDEADYYCQSYDTTQRV variable regionFGGGTKLTVL  388 SGSIADNY Clone PA14P1H05 - CDR-L1  389 EDNClone PA14P1H05 - CDR-L2  390 QSYDTTQRV Clone PA14P1H05 - CDR-L3  391QLQLQESGSRLVKPSQTLSLTCAVSGGSINSGGYSWSWIRQPPGKGLEWIClone PA12P3C09 - Heavy chainGNIYHGETTHYNPSLKSRVTISIDKSKNQFSLKLTSVTAADTAVYYCARA variable regionPLGNYYDTSGYLQPFDYWGPGALVTVSS  392 GGSINSGGYS Clone PA12P3C09 - CDR-H1 393 IYHGETT Clone PA12P3C09 - CDR-H2  394 ARAPLGNYYDTSGYLQPFDYClone PA12P3C09 - CDR-H3  395DIQMTQSPSSLSASVGDRVTITCRASQGIINDLGWYQQRPGRAPTRLIYAClone PA12P3C09 - light chainASSLQSGVPSRFSGSGSGTEFTLTINSLQPADFATYFCLQYNSYPPTFGQ variable regionGTKVEIK  396 QGIIND Clone PA12P3C09 - CDR-L1  149 AASClone PA12P3C09 - CDR-L2  397 LQYNSYPPT Clone PA12P3C09 - CDR-L3  398EVQLVESGGGVVRPGGSLRLSCAASGFIFRDHGMSWVRQAPGKGLEWVSGClone PA12P3C05 - Heavy chainINWNGANTGYADSVKGRSTISRDNAKNSLYLQMSSLRADDTALYHCVSHD variable regionYYYGLDVWGPGTTVIVSS  399 GFIFRDHG Clone PA12P3C05 - CDR-H1  400 INWNGANTClone PA12P3C05 - CDR-H2  401 VSHDYYYGLDV Clone PA12P3C05 - CDR-H3  402QSALTQPRSVSGSPGQSVTISCTGTSSDVGGDNYVSWYQQHPGKVPKLIIClone PA12P3C05 - light chainHDVSERPSGVPDRFSGSKSANTASLTISGLQADDEADYYCCSYAGTYTFG variable regionGGTRLTVL  403 SSDVGGDNY Clone PA12P3C05 - CDR-L1  404 DVSClone PA12P3C05 - CDR-L2  405 CSYAGTYT Clone PA12P3C05 - CDR-L3  406QVQLQESGPGLVKPSQTLSLTCTVSGGSISSGGYYWSWIRQLPGKGLEWIClone PA14P1G11 - Heavy chainGYIYYSGSTSYNPSLKSRVTISVDTSKNQLSLNLSSVTAADTAVYYCARG variable regionRRISISGVVTPLFDYWGQGTLVTVSS  199 GGSISSGGYY Clone PA14P1G11 - CDR-H1  200IYYSGST Clone PA14P1G11 - CDR-H2  201 ARGRRISISGVVTPLFDYClone PA14P1G11 - CDR-H3  407DIQMTQSPSSVSASVGDRVTITCRASQGISSWLAWYQQKPGKAPKLLIYAClone PA14P1G11 - light chainASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANSVPLTFGG variable regionGTKVEIK  203 QGISSW Clone PA14P1G11 - CDR-L1  149 AASClone PA14P1G11 - CDR-L2  204 QQANSVPLT Clone PA14P1G11 - CDR-L3  408QVQLVQSGAEVKKPGASVKVSCQASGYTFTRYDINWVRQATGQGLEWMGWClone PA12P3C01 - Heavy chainLNPKSGDTGYAQKFQGRVTMTRDTSISTAYMELTSLTSDDTAVYYCARGV variable regionDANHWGQGSLVTVSS  409 GYTFTRYD Clone PA12P3C01 - CDR-H1  410 LNPKSGDTClone PA12P3C01 - CDR-H2  411 ARGVDANH Clone PA12P3C01 - CDR-H3  412DIVVTQSPDSLAVSLGERATINCKSSQSIFDTSSNKNYLAWFRQRPGQPPClone PA12P3C01 - light chainQLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCHQYYSL variable regionPHAFGQGTKLEIK  413 QSIFDTSSNKNY Clone PA12P3C01 - CDR-L1   86 WASClone PA12P3C01 - CDR-L2  414 HQYYSLPHA Clone PA12P3C01 - CDR-L3  415EAQLVESGGGLVQPGGSLRLSCAASGFTFSSYYIHWVRQAPGKGLVWVSRClone PA16P1H09 - Heavy chainINSDGSSTRYADSVKGRFTISRDNAKNTLYLQMNSLRAEDTAVYFCARAS variable regionRTVYGTDSPLSNGMDVWGQGTKVVSS  416 GFTFSSYY Clone PA16P1H09 - CDR-H1  417INSDGSST Clone PA16P1H09 - CDR-H2  418 ARASRTVYGDSPLSNGMDVClone PA16P1H09 - CDR-H3  419SYELTQPPSVSVSPGQTASITCSGDKLGDKFACWYQQKPGHSPVLVIYQDClone PA16P1H09 - light chainDKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQAWDSSTHVVFGG variable regionGTKLTVL  420 KLGDKF Clone PA16P1H09 - CDR-L1  421 QDDClone PA16P1H09 - CDR-L2  422 QAWDSSTHVV Clone PA16P1H09 - CDR-L3  423EVQLVESGGGLVQPGGSLRLSCAASGFSFNTYNMNWVRQAPGKGLEWISDClone PA12P3D09 - Heavy chainITSSGSMRSYADAVKGRFTISRDNAKNSLHLQMNSLRVEDTAVYYCTRGW variable regionHDDLWSGYSYGLDVWGQGTTVTVSS  424 GFSFNTYN Clone PA12P3D09 - CDR-H1  425ITSSGSMR Clone PA12P3D09 - CDR-H2  426 TRGWHDDLWSGYSYGLDVClone PA12P3D09 - CDR-H3  427DIQMTQSPSSLSASVGDRVTITCRASQGIRNDLGWYQQRPGKAPRCLIYGClone PA12P3D09 - light chainASSLQSGVPSRFSGSGSGTEFTLTISNLQAEDFATYYCLQHKSYPLTFGP variable regionGTKVDIK  381 QGIRND Clone PA12P3D09 - CDR-L1   30 GASClone PA12P3D09 - CDR-L2  428 LQHKSYPLT Clone PA12P3D09 - CDR-L3  429QLLLLGPGPGVVRPSETLSLTCNVSGHSITDSPYYWGWIRQAPGKGLEWIClone PA13P3G04 - Heavy chainGHFYYSDYTYYNPSLKSRVNVSVDTSKNHLFLALTSVTAADTAVYYCARG variable regionFGGYDSPIWAIWGQGTLVTVSS  430 GHSITDSPYY Clone PA13P3G04 - CDR-H1  431FYYSDYT Clone PA13P3G04 - CDR-H2  432 ARGFGGYDSPIWAIClone PA13P3G04 - CDR-H3  433SHAVTQPPSVSVAPGQTASLTCAGDDIEENTVHWYQQKPGQAPVLVIYYTClone PA13P3G04 - light chainTDRPSAIPERFFGSKSGNTATLSIARVEAGDEADYYCQVSDRVFGGGTKL variable region TVL 434 DIEENT Clone PA13P3G04 - CDR-L1  435 YTT Clone PA13P3G04 - CDR-L2 436 QVSDRV Clone PA13P3G04 - CDR-L3  437EVQLVQSGGGLVKPGGSLRLSCAASGSTLTNYNINWVRQAPGKGLQWVSSClone PA11P1C03 - Heavy chainISGTRDYTYYADSVVGRFTISRDNAKNSVYLQMNSLRAEDTAVYYCARGR variable regionEVGGDYDSYDWGQGTLVTVSS  438 GSTLTNYN Clone PA11P1C03 - CDR-H1  439ISGTRDYT Clone PA11P1C03 - CDR-H2  440 ARGREVGGDYDSYDClone PA11P1C03 - CDR-H3  441DIQMTQSPSSLSASVGDRVTITCQASQDISTFLHWYQQKPGKAPSVLIYGClone PA11P1C03 - light chainASDLKTGVPSRFSGSGSGTHFTLTISSLQPEDIATYYCQQYDHLPLTFGG variable regionGTKVEIK  442 QDISTF Clone PA11P1C03 - CDR-L1   30 GASClone PA11P1C03 - CDR-L2  443 QQYDHLPLT Clone PA11P1C03 - CDR-L3  444EVQLVESGGGLIQPGGSLRLSCAASGFTVSSNYMSWVRQAPGKGLEWVSVClone PA11P1C01 - Heavy chainIYSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDSR variable regionLGWAYDAFDIWGQGTMVTVSS  445 GFTVSSNY Clone PA11P1C01 - CDR-H1  446IYSGGST Clone PA11P1C01 - CDR-H2  447 ARDSRLGWAYDAFDIClone PA11P1C01 - CDR-H3  448DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQClone PA11P1C01 - light chainLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQDGTFG variable regionQGTKVEIK   77 QSLLHSNGYNY Clone PA11P1C01 - CDR-L1   78 LGSClone PA11P1C01 - CDR-L2  449 MQDGT Clone PA11P1C01 - CDR-L3  450QVQLVQSGSELKKPGASVKASCKASGYTFSNYAVNWVRQAPGQGLEWMGWClone PA11P1C06 - Heavy chainINTKTGNPTYGQGFTGRFVFSLDTSVSTAYLQISSLKAEDTAVYYCARAA variable regionDYGEPYYYGMDVWGQGTTVTVSS  451 GYTFSNYA Clone PA11P1C06 - CDR-H1  452INTKTGNP Clone PA11P1C06 - CDR-H2  453 ARAADYGEPYYYGMDVClone PA11P1C06 - CDR-H3  454QSALTQPASVSGSPGQSITISCTGINSDVGSYNLVSWYQQHPGKAPKFMIClone PA11P1C06 - light chainYEGTKRPSGVSNRFSGSKSGHTASLTISGLQAEDEADYYCCSYAGTSTLV variable regionFGGGTKLTVL  455 NSDVGSYNL Clone PA11P1C06 - CDR-L1  456 EGTClone PA11P1C06 - CDR-L2  457 CSYAGTSTLV Clone PA11P1C06 - CDR-L3  458EVQLVESGGGLVQPGGSLRLSCAASGFTFRNYWMNWVRQAPGKGLVWVSRClone PA13P1H03 - Heavy chainINSEGSSTSYADPVKGRFTISRDNAKDTLYLQMDSLRAEDSAVYYCARIF variable regionNGYIHVGRDYWGQGTRVTVSS  459 GFTFRNYW Clone PA13P1H03 - CDR-H1  460INSEGSST Clone PA13P1H03 - CDR-H2  461 ARIFNGYIHVGRDYClone PA13P1H03 - CDR-H3  462DIQMTQSPSTLSASIGDRVTITCRASESISNWLAWFQQKPGKAPKLLIYKClone PA13P1H03 - light chainASNLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQYNSNSQTFGQ variable regionGTKLDLK  463 ESISNW Clone PA13P1H03 - CDR-L1  196 KASClone PA13P1H03 - CDR-L2  464 QQYNSNSQT Clone PA13P1H03 - CDR-L3  465QVQLQESGPGLVKPSETLSLTCTVSGGSISSYYWNWIRQPPGKGLEWIGYClone PA11P1C04 - Heavy chainIYYSGSTNYNPSLKSRATISVDTSKNQFSLKLSSVTAADTAVYYCARANL variable regionFGVALRRVLGPFDYWGQGTLVTVSS  466 GGSISSYY Clone PA11P1C04 - CDR-H1  200IYYSGST Clone PA11P1C04 - CDR-H2  467 ARANLFGVALRRVLGPFDYClone PA11P1C04 - CDR-H3  468DIQMTQSPSSLSASVGDRVTIACRASQSIANYLNWYQQKPGKAPKLLIYAClone PA11P1C04 - light chainASNLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPYTFGQ variable regionGTKLEIK  469 QSIANY Clone PA11P1C04 - CDR-L1  149 AASClone PA11P1C04 - CDR-L2  470 QQSYSTPYT Clone PA11P1C04 - CDR-L3  471QVQLVESGGGVVQPGRSLRLSCAASGFSFRSYGMHWVRQAPGKGLEWVAVClone PA14P1D10 - Heavy chainISYDGSNKYYVDSVKGRFTISRDNSKNTLYVQMNSLTDEDTAVYYCARDR variable regionGVTTRQFSYYYYGMDVWGQGTTVTVSS  472 GFSFRSYG Clone PA14P1D10 - CDR-H1   59ISYDGSNK Clone PA14P1D10 - CDR-H2  473 ARDRGVTTRQFSYYYYGMDVClone PA14P1D10 - CDR-H3  474AIRMTQSPSSFSASTGDRVTITCRASQSITSYLAWYQQKPGKAPKLLIYAClone PA14P1D10 - light chainASTLQSGLPSRFSGSGSGTDFTLTISGLQSEDFATYYCQQYYNYPQTFGQ variable regionGTRVEIK  475 QSITSY Clone PA14P1D10 - CDR-L1  149 AASClone PA14P1D10 - CDR-L2  476 QQYYNYPQT Clone PA14P1D10 - CDR-L3  477EVQLLESGGQLVQPGGSLRLSCGAFGFTFGDAAMTWVRQAPGKGLEWVSTClone PA14P1C10 - Heavy chainISGRGDETFSADSVKGRFTISRDNFKNMLYVQMNSLRAEDTATYYCARLG variable regionHLRGWFDSWGQGTLVTVSS  478 GFTFGDAA Clone PA14P1C10 - CDR-H1  479 ISGRGDETClone PA14P1C10 - CDR-H2  480 ARLGHLRGWFDS Clone PA14P1C10 - CDR-H3  481EIVMTQSPATLSVSPGERVTLSCRASQSVSSNLAWYQQKPGQAPRLLIYGClone PA14P1C10 - light chainASTRATGIPAGFSGSGSGTEFTLTISSLQSEDFAVYYCQQYNNWPRTFGQ variable regionGTKVEIK  482 QSVSSN Clone PA14P1C10 - CDR-L1   30 GASClone PA14P1C10 - CDR-L2  252 QQYNNWPRT Clone PA14P1C10 - CDR-L3  483EVQLVESGGGLVKPGGSLRLSCAASGFTFSDAWMTWVRQAPGKGLEWVGRClone PA11P1C08 - Heavy chainIKSKTDGGTTDYGAPVKGRFSISRDDSKNTLYLHMNSLKTEDTAVYYCTT variable regionKSPNSNWFPFYYYYYMDVWGKGTTVTVSS  484 GFTFSDAW Clone PA11P1C08 - CDR-H1 485 IKSKTDGGTT Clone PA11P1C08 - CDR-H2  486 TTKSPNSNWFPFYYYYYMDVClone PA11P1C08 - CDR-H3  487QSALTQPRSVSGSPGQSVTISCTGTSSDVGGYNFVSWYQQHPGKAPQLMIClone PA11P1C08 - light chainYDVTKRPSGVPDRFSGSKSGNTASLTISGLQAEDEGDYYCYSYAASSLYV variable regionFGTGTKVTVL  488 SSDVGGYNF Clone PA11P1C08 - CDR-L1  141 DVTClone PA11P1C08 - CDR-L2  489 YSYAASSLYV Clone PA11P1C08 - CDR-L3  490QLQLQESGPGLVKPSETLSLICTVSGGAITSSTFYWAWIRQPPGRGLEWIClone PA14P3F10 - Heavy chainGSMYYSGSTYYNLSLKSRVIISVNTSKNQFSLTLTSATATDMAVYYCVRH variable regionTLHDYGSGSFPDYSYGMDVWGQGTTVTVSS  491 GGAITSSTFY Clone PA14P3F10 - CDR-H1 492 MYYSGST Clone PA14P3F10 - CDR-H2  493 VRHTLHDYGSGSFPDYSYGMDVClone PA14P3F10 - CDR-H3  494EIVLTQSPATLSLFPGERGTLSCRASQSVSSHLIWYQQKPGQAPRVLIFDClone PA14P3F10 - light chainATNRATGIPARFSGSGSGTDFTLTISNLEPEDYGVYYCQQRSNWPLTFGG variable regionGTKVEIK  495 QSVSSH Clone PA14P3F10 - CDR-L1  265 DATClone PA14P3F10 - CDR-L2  496 QQRSNWPLT Clone PA14P3F10 - CDR-L3  497EVQLLESGGGLVQPGGSLKLSCVASGFTFSSYAMMWVRQAPGKGLEWISSClone PA14P1D11 - Heavy chainISSSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRVEDTAVYYCAKSH variable regionCSTTSCPRAFYYYGMDVWGQGTTVTVSS  113 GFTFSSYA Clone PA14P1D11 - CDR-H1  498ISSSGGST Clone PA14P1D11 - CDR-H2  499 AKSHCSTTSCPRAFYYYGMDVClone PA14P1D11 - CDR-H3  500DIQMTQSPSSLSASVGDRVTITCRASQTITTYLNWYQQKPGKAPKLLIYAClone PA14P1D11 - light chainASSLQSGVPSRFSGSGSGTDFTLSLSSLQPEDSATYYCQQSYSTLGAFGG variable regionGTKVEIK  501 QTITTY Clone PA14P1D11 - CDR-L1  149 AASClone PA14P1D11 - CDR-L2  502 QQSYSTLGA Clone PA14P1D11 - CDR-L3  503QVHLQESGPGLVKPSGTLSLTCTVSGGSISTYYWSWIRQPPGKGLEWIGYClone PA14P3F02 - Heavy chainIYYGGTTNYNPSLKSRVTISVDTSKNQFSLRLRSVTAADTAVYYCAREID variable regionSRMDRWGQGTLVTVSS  504 GGSISTYY Clone PA14P3F02 - CDR-H1  505 IYYGGTTClone PA14P3F02 - CDR-H2  506 AREIDSRMDR Clone PA14P3F02 - CDR-H3  507SYALTQPPSVSVAPGKTARITCGGDNIGSKTVHWYHQKPGQAPVLVIYYDClone PA14P3F02 - light chainSNRPSGISERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSNSDHRIFG variable regionGGTKLTVL  508 NIGSKT Clone PA14P3F02 - CDR-L1  509 YDSClone PA14P3F02 - CDR-L2  510 QVWDSNSDHRI Clone PA14P3F02 - CDR-L3  511QVQLVQSGAEVRKPGSSVKVSCKASGGTFSNNPITWVRQAPGQGLEWMGWClone PA12P1G11 - Heavy chainIIPIFNTTNYAQKFQGRVTITADESTSTAYMELSSLKSEDTALFYCARDR variable regionAHAYCNNGVCYTTDAFDVWGQGTLVTVSS  512 GGTFSNNP Clone PA12P1G11 - CDR-H1 513 IIPIFNTT Clone PA12P1G11 - CDR-H2  514 ARDRAHAYCNNGVCYTTDAFDVClone PA12P1G11 - CDR-H3  515ETVLTQSPATLSLSPGERATLSCRASQSVGRYLAWYQHKPGQAPRLLIYDClone PA12P1G11 - light chainASNRATGIPARFSGSGSGTDFTLTISSLEPEDSAVYYCQQGTDWLTFGGG variable regionTKVEIK  516 QSVGRY Clone PA12P1G11 - CDR-L1   94 DASClone PA12P1G11 - CDR-L2  517 QQGTDWLT Clone PA12P1G11 - CDR-L3  518QVQLVESGGGLVKPGGSLRLSCAASGITFSDNYMTWIRQAPGKGLEWVSYClone PA13P1E06 - Heavy chainISSSGTNIFYADSLKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARTL variable regionMTGSSLYFDYWGQGTQVTVSS  519 GITFSDNY Clone PA13P1E06 - CDR-H1  520ISSSGTNI Clone PA13P1E06 - CDR-H2  521 ARTLMTGSSLYFDYClone PA13P1E06 - CDR-H3  522SYELTQPPSVSVSPGQTARITCSGDALPKQYAYWYQQKPGQAPVLVIYKDClone PA13P1E06 - light chainSERPSGIPERFSGSSSGTTVTLAISGVQAEDEADYYCQSADIRVTESVLF variable regionGGGTKLTVL  523 ALPKQY Clone PA13P1E06 - CDR-L1  524 KDSClone PA13P1E06 - CDR-L2  525 QSADIRVTESVL Clone PA13P1E06 - CDR-L3  526EVHLLESGGHLVQPGGSLRLACAVSGFTFSDSAMTWVRQAPGKGLEWVSTClone PA14P1C12 - Heavy chainISGRGDETFFADSVKGRFSIFRDNSNSVLYVQMNSLRAEDTATYYCARYG variable regionHHKGWFDSWGQGTLVTVSS  527 GFTFSDSA Clone PA14P1C12 - CDR-H1  479 ISGRGDETClone PA14P1C12 - CDR-H2  528 ARYGHHKGWFDS Clone PA14P1C12 - CDR-H3  529EIVMTQSPATLSVSPGERATLSCRASQSVSSNLAWYQQKPGQAPRLLIYGClone PA14P1C12 - light chainASTRAIGIPAGFSGSGSGTEFTLTISSLQSEDSAVYYCQQYNNWPRTFGQ variable regionGTKVEIK  482 QSVSSN Clone PA14P1C12 - CDR-L1   30 GASClone PA14P1C12 - CDR-L2  252 QQYNNWPRT Clone PA14P1C12 - CDR-L3  530QVQLVQSGTEVKKPGASVKVSCKASGYTFSSFGITWVRQAPGQGLEWMGWClone PA11P1D07 - Heavy chainISAYNGNTKYAQAVQGRVTLTTDTSTTTAYMELRSLRSNDTAVYFCAREG variable regionIEHLVVEGRGPGGDCWGQGTLVIVSS  531 GYTFSSFG Clone PA11P1D07 - CDR-H1  532ISAYNGNT Clone PA11P1D07 - CDR-H2  533 AREGIEHLVVEGRGPGGDCClone PA11P1D07 - CDR-H3  534SYELTQPPSVSVSPGQTARITCSGDALPKEYTSWYQQKSGQAPVLVIYEDClone PA11P1D07 - light chainIKRPSGIPERFSGSSSGTMASLTISGAQVDDEADYYCYSTDTSGDHKVFG variable regionGGTKLTVL  535 ALPKEY Clone PA11P1D07 - CDR-L1  536 EDIClone PA11P1D07 - CDR-L2  537 YSTDTSGDHKV Clone PA11P1D07 - CDR-L3  538QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYAMHWVRQAPGKGREWVAAClone PA12P4D02 - Heavy chainLSYDGSSTYYADSVKGRLTISRDNSKNTLYLQMNSLRAEDTAVYFCTRVP variable regionYGEGRAANDYWGQGTLVTVSS  113 GFTFSSYA Clone PA12P4D02 - CDR-H1  539LSYDGSST Clone PA12P4D02 - CDR-H2  540 TRVPYGEGRAANDYClone PA12P4D02 - CDR-H3  541DIQMTQSPSTLSASVGDRVTITCRASQSIGSWLAWYQQKPGKAPKLLIYKClone PA12P4D02 - light chainASNIESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQHYNTYSRSFGG variable regionGTEVAIK  542 QSIGSW Clone PA12P4D02 - CDR-L1  196 KASClone PA12P4D02 - CDR-L2  543 QHYNTYSRS Clone PA12P4D02 - CDR-L3  544QVQLVQSGAEVKTPGSSVKVSCTASGDSFSRYAINWVRQAPGQGLEWVGKClone PA15P1C03 - heavy chainIVPVFGAASYAQKFQGRVTITADESTSTVYMELSSLRSEDTAVYYCARGI variable regionVKLSTMPPVYWGQGTLVTVSS  545 GDSFSRYA Clone PA15P1C03 - CDR-H1  546IVPVFGAA Clone PA15P1C03 - CDR-H2  547 ARGIVKLSTMPPVYClone PA15P1C03 - CDR-H3  548DIVMTQTPLSLSVTPGQPASISCKSSQSLLHSDGKTYLYWYLQKPGQSPQClone PA15P1C03 - light chainLLISEVSSRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQGIHHL variable regionTFGPGTKVDIK  549 QSLLHSDGKTY Clone PA15P1C03 - CDR-L1  550 EVSClone PA15P1C03 - CDR-L2  551 MQGIHHLT Clone PA15P1C03 - CDR-L3  552QVQLQESGPGLVKPSETLSLTCSVSGGSVSDSAYYWSWIRQPPGGGLEFIClone PA12P4G06 - heavy chainGYVYNSGSTNYNPSLKSRVTISVDTSKNQFSLSLSSLTAADTAVYYCARY variable regionCSSTSCYVRSSDVNWFDPWGQGTLVIVSS  553 GGSVSDSAYY Clone PA12P4G06 - CDR-H1 554 VYNSGST Clone PA12P4G06 - CDR-H2  555 ARYCSSTSCYVRSSDVNWFDPClone PA12P4G06 - CDR-H3  556EIVLTQSPGTLSSSPGESATLSCRASQSLGTYLAWYQQKPGQAPRLLIYDClone PA12P4G06 - light chainASKRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCHQRSHWLTFGGG variable regionTKVEIK  557 QSLGTY Clone PA12P4G06 - CDR-L1   94 DASClone PA12P4G06 - CDR-L2  558 HQRSHWLT Clone PA12P4G06 - CDR-L3  559EAQLLESGGGLVQPGGSLRLSCAASGFNFSNYAMTWVRQAPGKGLEWVSAClone PA16P1B09 - heavy chainISSGGGTTYYADSVKGRFTISRDNSKNTVYLQMNSLKDADSALYYCAKPG variable regionRAVVVRLSYFDSWGQGTLVTVSS  560 GFNFSNYA Clone PA16P1B09 - CDR-H1  561ISSGGGTT Clone PA16P1B09 - CDR-H2  562 AKPGRAVVVRLSYFDSClone PA16P1B09 - CDR-H3  563QSVLTQPPSVSAAPGQKVSISCSGSGSNIANHYVSWYQHLPGTAPKLLIYClone PA16P1B09 - light chainDNNKRPSGIPDRFSGSKSGTSATLGITGLQTGDEADYYCGTWDSSLTVVV variable regionFGGGTKLTVL  564 GSNIANHY Clone PA16P1B09 - CDR-L1  110 DNNClone PA16P1B09 - CDR-L2  565 GTWDSSLTVVV Clone PA16P1B09 - CDR-L3  566QITLKESGPTLVKPTETLTLTCTFSGFSLTTSGVAVGWVRQPPGKALEWLClone PA12P4G03 - heavy chainALIYWDDDERYTPSLKSRLTITKDTSKSQVVLTMTNMDPVDTATYFCVHC variable regionEGPDILLVPAAYFFDFWGQGTLVTVSS  567 GFSLTTSGVA Clone PA12P4G03 - CDR-H1 568 IYWDDDE Clone PA12P4G03 - CDR-H2  569 VHCEGPDILLVPAAYFFDFClone PA12P4G03 - CDR-H3  570EIVLTQSPGTLSLSPGDRATLSCRASQSVSRRYLAWYQQSPGQAPRLLISClone PA12P4G03 - light chainGASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAMYYCQQYGSSTGTFG variable regionQGTKVEMK  571 QSVSRRY Clone PA12P4G03 - CDR-L1   30 GASClone PA12P4G03 - CDR-L2  572 QQYGSSTGT Clone PA12P4G03 - CDR-L3  573QVQLQESGPGLVKPSQTLSLTCTVSGGSISSSGYYWSWIRQHPGKGLEWIClone PA11P1F10 - heavy chainGYINYIGGTYYNPSLRSRVTMSVDTSKNQFSLRLSSVSAADTAVYYCAST variable regionHSYGDYSRDYYYGVDVWGQGTTVTISS  574 GGSISSSGYY Clone PA11P1F10 - CDR-H1 575 INYIGGT Clone PA11P1F10 - CDR-H2  576 ASTHSYGDYSRDYYYGVDVClone PA11P1F10 - CDR-H3  577EIVLTQSPATLSLSPGDRATLSCRTSQSVSSSYLAWYQQKPGQAPRLLIYClone PA11P1F10 - light chainAASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQCAGSPFTFG variable regionPGTKVDLK   62 QSVSSSY Clone PA11P1F10 - CDR-L1  149 AASClone PA11P1F10 - CDR-L2  578 QQCAGSPFT Clone PA11P1F10 - CDR-L3  579QVQLVESGGGVVQPGRSLRLSCAASGFTFSDYAMHWVRQAPGKGLEWVAVClone PA12P1G02 - heavy chainISYDGNHRYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARHP variable regionGLSIAVAGPFDYWGQGTLVTVSS  580 GFTFSDYA Clone PA12P1G02 - CDR-H1  581ISYDGNHR Clone PA12P1G02 - CDR-H2  582 ARHPGLSIAVAGPFDYClone PA12P1G02 - CDR-H3  583EIVMTQSPATLSVSPGERATLSCGASQSVSSNLAWYQQKPGQAPRLLFYGClone PA12P1G02 - light chainASTRATGIPARFSGSGSGTEFTLTISSLQSEDFALYYCQQYNNWPWTFGQ variable regionGTKVDIK  482 QSVSSN Clone PA12P1G02 - CDR-L1   30 GASClone PA12P1G02 - CDR-L2  584 QQYNNWPWT Clone PA12P1G02 - CDR-L3  585QVQLVQSGAEVKKPGASVKVSCKASGYTFNRDGITWVRQAPGQGLEWMGWClone PA16P1E12 - heavy chainISANNDFTDYAQKFQGRLTMTTDTSTNTAYMELRSLRSDDTAVYYCARQV variable regionITVLQYSYGMDVWGQGTTVTVSS  586 GYTFNRDG Clone PA16P1E12 - CDR-H1  587ISANNDFT Clone PA16P1E12 - CDR-H2  588 ARQVITVLQYSYGMDVClone PA16P1E12 - CDR-H3  589DIQMTQFPSSLSASVGDRVTITCRASQSISRYLNWYQQTPGKAPKLLIYGClone PA16P1E12 - light chainASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSDTAPLTFGG variable regionGTRVEIK  590 QSISRY Clone PA16P1E12 - CDR-L1   30 GASClone PA16P1E12 - CDR-L2  591 QQSDTAPLT Clone PA16P1E12 - CDR-L3  415EAQLVESGGGLVQPGGSLRLSCAASGFTFSSYYIHWVRQAPGKGLVWVSRClone PA16P1E11 - heavy chainINSDGSSTRYADSVKGRFTISRDNAKNTLYLQMNSLRAEDTAVYFCARAS variable regionRTVYGDSPLSNGMDVWGQGTKVTVSS  416 GFTFSSYY Clone PA16P1E11 - CDR-H1  417INSDGSST Clone PA16P1E11 - CDR-H2  418 ARASRTVYGDSPLSNGMDVClone PA16P1E11 - CDR-H3  419SYELTQPPSVSVSPGQTASITCSGDKLGDKFACWYQQKPGHSPVLVIYQDClone PA16P1E11 - light chainDKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQAWDSSTHVVFGG variable regionGTKLTVL  420 KLGDKF Clone PA16P1E11 - CDR-L1  421 QDDClone PA16P1E11 - CDR-L2  422 QAWDSSTHVV Clone PA16P1E11 - CDR-L3  592QVQLVESGGGLVTPGGSLRLSCTVSGFTLSDYYMSWIRQAPGKGLDWLSYClone PA12P3E11 - heavy chainISGSGDNKNYADSVRGRFTISRDNSKNSLYLQMNSLRAEDTAVYYCAREF variable regionPSGGYSPGVVLWGQGTLVTVSS  593 GFTLSDYY Clone PA12P3E11 - CDR-H1  594ISGSGDNK Clone PA12P3E11 - CDR-H2  595 AREFPSGGYSPGVVLClone PA12P3E11 - CDR-H3  596NFVLTQPHSVSESPGKTVTISCARSSGSIAGSFVQWYQQRPGSSPTTVIYClone PA12P3E11 - light chainEDTRRPSGVPDRFSGSIDSSSNSASLTISGLKTEDEADYYCQSYDSTNPW variable regionVFGGGTKLTVL  597 SGSIAGSF Clone PA12P3E11 - CDR-L1  598 EDTClone PA12P3E11 - CDR-L2  599 QSYDSTNPWV Clone PA12P3E11 - CDR-L3  600EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSVClone PA14P1C06 - heavy chainIYSGGSRTYYADSAKGRFTISRDNSKNMLYLQMNSLRAEDTAVYYCTKND variable regionYDSSGYFDFDNWGQGTLVTVSS  113 GFTFSSYA Clone PA14P1C06 - CDR-H1  114IYSGGSRT Clone PA14P1C06 - CDR-H2  601 TKNDYDSSGYFDFDNClone PA14P1C06 - CDR-H3  602QSALTQPASVSGSPGQSITISCTGTSSDVGGYNYVSWYQQHPGKAPKLMIClone PA14P1C06 - light chainYDVSNRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYHCSSYTSSSTWV variable regionFGGGTKLTVL  603 SSDVGGYNY Clone PA14P1C06 - CDR-L1  404 DVSClone PA14P1C06 - CDR-L2  604 SSYTSSSTWV Clone PA14P1C06 - CDR-L3  605EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSVClone PA14P1C07 - heavy chainVYSGGSGTYYADSVKGRFTISRDDSTNTLYLQMNSLRAEDTAVYYCAKDR variable regionDSFGELDLDSWGQGTLVSVSS  113 GFTFSSYA Clone PA14P1C07 - CDR-H1  606VYSGGSGT Clone PA14P1C07 - CDR-H2  607 AKDRDSFGELDLDSClone PA14P1C07 - CDR-H3  608DIVMTQSPDSLAVSLGERATINCKSSQSVLYSSKNKNYLAWYQQRPGQPPClone PA14P1C07 - light chainKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCLQYYNI variable regionPRTFGQGTKLEIK  609 QSVLYSSKNKNY Clone PA14P1C07 - CDR-L1   86 WASClone PA14P1C07 - CDR-L2  610 LQYYNIPRT Clone PA14P1C07 - CDR-L3  611EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYALSWVRQPPGKGLEWVSVClone PA14P1C04 - heavy chainIYSGGSRTYYADAAKGRFTISRDNSKNMLYLQMNSLRAEDTAVYYCTKHD variable regionYDSSGYFDFDNWGQGTLVTVSS  113 GFTFSSYA Clone PA14P1C04 - CDR-H1  114IYSGGSRT Clone PA14P1C04 - CDR-H2  612 TKHDYDSSGYFDFDNClone PA14P1C04 - CDR-H3  602QSALTQPASVSGSPGQSITISCTGTSSDVGGYNYVSWYQQHPGKAPKLMIClone PA14P1C04 - light chainYDVSNRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYHCSSYTSSSTWV variable regionFGGGTKLTVL  603 SSDVGGYNY Clone PA14P1C04 - CDR-L1  404 DVSClone PA14P1C04 - CDR-L2  604 SSYTSSSTWV Clone PA14P1C04 - CDR-L3  613EVQLVESGGGVARPGGSLRLSCAASGFTFDDYGMSWVRQAPGKGLEWVSGClone PA14P1C02 - heavy chainYINWNGGSTNADSVKGRFIISRDNGKNSLYLQMNSLRAEDTAFYHCARRG variable regionNFYYYGMDVWGQGTTVTVSS  614 GFTFDDYG Clone PA14P1C02 - CDR-H1  615INWNGGST Clone PA14P1C02 - CDR-H2  616 ARRGNFYYYGMDVClone PA14P1C02 - CDR-H3  617DIQMTQSPSSVSASVGDRVTITCRASQGNSTWLAWYQQKPGKAPELLIFDClone PA14P1C02 - light chainASNLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQAQRFPLTFGG variable regionGTKVEIK  618 QGNSTW Clone PA14P1C02 - CDR-L1   94 DASClone PA14P1C02 - CDR-L2  619 QQAQRFPLT Clone PA14P1C02 - CDR-L3  620QVQLVQSGSELRRPGASVKVSCKTSGYAFTHFAMNWLRQAPGQGLEWLGWClone PA11P1E01 - heavy chainINTHSGNPTYAQGFTGRIVFSLDTSAGTAYLEISSLKAEDTAVYYCARER variable regionYFDFWGQGTLVAVSS  621 GYAFTHFA Clone PA11P1E01 - CDR-H1  176 INTHSGNPClone PA11P1E01 - CDR-H2  177 ARERYFDF Clone PA11P1E01 - CDR-H3  622QSVLTQPPSASGTPGQRVTISCSGTNSNIGKNFLYWYQQLPGTAPKLLIFClone PA11P1E01 - light chainSSNQRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDNLSGWV variable regionFGGGTKVTVL  623 NSNIGKNF Clone PA11P1E01 - CDR-L1  180 SSNClone PA11P1E01 - CDR-L2  624 AAWDDNLSGWV Clone PA11P1E01 - CDR-L3  625QVQLQESGPGLVKPSEALSLTCSVSDGSVSSGSYYWTWIRQPPGKGLEWIClone PA11P1E08 - heavy chainGCIHYSGRTNYNPSLKSRVTISIDTSKNQFSLQLSSVTAVDTAVYYCARD variable regionRGEYDFWRVRYYGMDVWGQGTTVTVSS  626 DGSVSSGSYY Clone PA11P1E08 - CDR-H1 627 IHYSGRT Clone PA11P1E08 - CDR-H2  628 ARDRGEYDFWRVRYYGMDVClone PA11P1E08 - CDR-H3  629QSALTQPASVSGSPGQSITISCTGTSSDVGDYNYVSWYQQHPGKAPKLLIClone PA11P1E08 - light chainYDFSNRPSGVSDRFSGSKSGNTASLTISGLRAEDESDYYCTSYTNTNTRL variable regionFGGGTKLTVL  630 SSDVGDYNY Clone PA11P1E08 - CDR-L1  631 DFSClone PA11P1E08 - CDR-L2  632 TSYTNTNTRL Clone PA11P1E08 - CDR-L3  633QVQLVQSGAEVKKPGASVKVSCKASGYTFISYGLHWVRQAPGQRPEWMGWClone PA14P1C08 - heavy chainINAGNGNRKYSERFQARVTFTRDTSATTAYMELSSLRSEDTAVYYCARDR variable regionLTAAAHFDYWGQGTQVTVSS  634 GYTFISYG Clone PA14P1C08 - CDR-H1  635INAGNGNR Clone PA14P1C08 - CDR-H2  636 ARDRLTAAAHFDYClone PA14P1C08 - CDR-H3  637EIVMTQSPATLSVSLGERATLSCRASQSVSSDLAWYQQKPGQAPRLLMYGClone PA14P1C08 - light chainASTRATGFPARFTGSGSGPEFTLTISSLQSEDFAVYYCQQYNNWPFTFGG variable regionGTKVEIK  638 QSVSSD Clone PA14P1C08 - CDR-L1   30 GASClone PA14P1C08 - CDR-L2  639 QQYNNWPFT Clone PA14P1C08 - CDR-L3  640QITLKESGPTLVKPTQTLTLTCTFSGFSLTSSAVGVGWIRQPPGKALEWLClone PA14P1H12 - heavy chainALIYGDDDKRYSPSLKRRLTITKDTSKNQVVLTMTDVDPVDTATYYCAHR variable regionRLTIPLLMVAADAFDIWGPGTMVIVSS  641 GFSLTSSAVG Clone PA14P1H12 - CDR-H1 642 IYGDDDK Clone PA14P1H12 - CDR-H2  643 AHRRLTIPLLMVAADAFDIClone PA14P1H12 - CDR-H3  644DIQMTQSPSTLSASVGDRVTITCRASQSVSRWLAWYQQKPGKAPKLLIYRClone PA14P1H12 - light chainASSLQSGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQYSSFHTFGQG variable regionTKLEIK  645 QSVSRW Clone PA14P1H12 - CDR-L1  646 RASClone PA14P1H12 - CDR-L2  647 QQYSSFHT Clone PA14P1H12 - CDR-L3  648EVQLVESGGGLVQPGGSLRLSCAASGFIFSTYSMNWVRQAPGKGLEWVSYClone PA14P1H11 - heavy chainISSSSNTIYYADSVKGRFTISRDNAKNSLYLQMNSLRDADTAVYYCARDG variable regionGRSGYFDDYWGQGTLVTVSS  649 GFIFSTYS Clone PA14P1H11 - CDR-H1  650ISSSSNTI Clone PA14P1H11 - CDR-H2  651 ARDGGRSGYFDDYClone PA14P1H11 - CDR-H3  652QLVLTQSPSASASLGASVKLTCTLSNGHINYAIAWHQQQPDKGPRYLLNLClone PA14P1H11 - light chainPKSDGSHSKGDGIDRFSGSSSGAERYLTISGLQSEDEADYYCQTWGTGIQ variable regionVFGGGTKLTVL  653 NGHINYA Clone PA14P1H11 - CDR-L1  654 LKSDGSHClone PA14P1H11 - CDR-L2  655 QTWGTGIQV Clone PA14P1H11 - CDR-L3  656QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEClone PA14P1D02 - heavy chainINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGRG variable regionVWFGELFPFDYWGQGTLGTVSS  657 GGSFSGYY Clone PA14P1D02 - CDR-H1  658INHSGST Clone PA14P1D02 - CDR-H2  659 ARGRGVWFGELFPFDYClone PA14P1D02 - CDR-H3  660QGGLTQPPSVSKGLRQTATLTCTGNSNNVGNQGAAWLQQHQGHPPKLLSYClone PA14P1D02 - light chainRNNNRPSGISERFSASRSGNTASLTITGLQPEDEADYYCSAWDSSLSAVV variable regionFGGGTKLTVL  661 SNNVGNQG Clone PA14P1D02 - CDR-L1  662 RNNClone PA14P1D02 - CDR-L2  663 SAWDSSLSAVV Clone PA14P1D02 - CDR-L3  664QVQLQQWGAGLLKPSETLSLNCTVYHGSLSTSYWSWIRQPPGRGLEWIGEClone PA11P1F03 - heavy chainINDSGATNYNPSLKSRVIISVDTSKDQFSLKLTSVTAADTAMYYCARAPL variable regionLWVGESFFYYFDSWGQGILVTVSS  665 HGSLSTSY Clone PA11P1F03 - CDR-H1  666INDSGAT Clone PA11P1F03 - CDR-H2  667 ARAPLLWVGESFFYYFDSClone PA11P1F03 - CDR-H3  668DIQMTQSPSSLSASVGDRVSITCRAGQSIDTYLNWYQHKPGKAPDLLIYTClone PA11P1F03 - light chainGTSTLHSGVPSRFSSGSGTDFTLTITSLQPEDFAIYYCQQSYKSPYTFGQ variable regionGTKVEIK  669 QSIDTY Clone PA11P1F03 - CDR-L1  670 TTSClone PA11P1F03 - CDR-L2  671 QQSYKSPYT Clone PA11P1F03 - CDR-L3  672QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIClone PA11P1F02 - heavy chainINPSGGTTSSAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTALYYCARDR variable regionEQKVGGAPLHWGQGTLVTVSS   89 GYTFTSYY Clone PA11P1F02 - CDR-H1  673INPSGGTT Clone PA11P1F02 - CDR-H2  674 ARDREQKVGGAPLHClone PA11P1F02 - CDR-H3  675DIQMTQSPSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYDClone PA11P1F02 - light chainASNLETGVPSRFSGSGSGTDFSFTISSLQPEDIATYYCQQYDNFALTFGG variable regionGTKVEIK  374 QDISNY Clone PA11P1F02 - CDR-L1   94 DASClone PA11P1F02 - CDR-L2  676 QQYDNFALT Clone PA11P1F02 - CDR-L3  677EVQLLESGGGLVQPGVSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSVClone PA14P1D07 - heavy chainIYSGGNIIYYADSVKGRFTISRDNSKNTLYLQIDNLRAEDTALYYCAKHD variable regionYDSSGYFDFDYWGHGTLVTVSS  113 GFTFSSYA Clone PA14P1D07 - CDR-H1  678IYSGGNII Clone PA14P1D07 - CDR-H2  679 AKHDYDSSGYFDFDYClone PA14P1D07 - CDR-H3  680QSALTQPASVSGSPGQSITISCTGTSRDVGGYNYVSWYQQHPGKAPKLMIClone PA14P1D07 - light chainYDVNNRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYFCCSYTSSSTWV variable regionFGGGTKLTVL  681 SRDVGGYNY Clone PA14P1D07 - CDR-L1  682 DVNClone PA14P1D07 - CDR-L2  683 CSYTSSSTWV Clone PA14P1D07 - CDR-L3  684QVQLQESGPGLVKPSGTLSLTCAVSGASISNSAWWNWVRQPPRGGLEWVGClone PA14P1D09 - heavy chainEIYPSGSTNYTPSLKSRATILLDESRNEFSLKLNSVTAADTAVYYCARGR variable regionLEDCNGGVCYFFDNWGQGILVSVSS  685 GASISNSAW Clone PA14P1D09 - CDR-H1  686IYPSGST Clone PA14P1D09 - CDR-H2  687 ARGRLEDCNGGVCYFFDNClone PA14P1D09 - CDR-H3  688DIEMTQSPSTLSASVGDRVTITCRANYGIGAWLAWYQQKPGKAPKLLIYKClone PA14P1D09 - light chainASTLESGVPLRFSGSGSGTEFTLSISGLQPDDFATYYCHQYSTYPITFGQ variable regionGTRLEIK  689 YGIGAW Clone PA14P1D09 - CDR-L1  196 KASClone PA14P1D09 - CDR-L2  690 HQYSTYPIT Clone PA14P1D09 - CDR-L3  691QVQLVESGGGVVQPGRSLTLSCAASGFNFKTYGMHWVRQAPGKGLEWVAVClone PA14P3H08 - heavy chainIYHDGNDKFYADSVKGRFTISRDNSKNTLYVQMSSLRADDTAIYYCAKGI variable regionFSSGYHYGMDVWGQGTAVIVSS  692 GFNFKTYG Clone PA14P3H08 - CDR-H1  693IYHDGNDK Clone PA14P3H08 - CDR-H2  694 AKGIFSSGYHYGMDVClone PA14P3H08 - CDR-H3  695DIQMTQSPSSLSASLGDSVTITCLASQGIKEFLSWFQQKPGQAPKLLIYDClone PA14P3H08 - light chainSASSSHSGVPSRFSGGSATHFTLTISSLQPDDIATYYCQQYHQVPLTFGQ variable regionGTRLEIK  696 QGIKEF Clone PA14P3H08 - CDR-L1   94 DASClone PA14P3H08 - CDR-L2  697 QQYHQVPLT Clone PA14P3H08 - CDR-L3  190QVQLQQWGAGLLKPSETLSLTCVVSGGSFSTHYWNWIRQSPGKGLEWIGEClone PA15P1E01 - heavy chainINHSGNTNYNPSLTGRATISVATSKTQFSLRLNSVTAADTAVYFCARGPR variable regionLRYTAGRPLFDTWGQGTLVTVSS  191 GGSFSTHY Clone PA15P1E01 - CDR-H1  192INHSGNT Clone PA15P1E01 - CDR-H2  193 ARGPRLRYTAGRPLFDTClone PA15P1E01 - CDR-H3  194DIQMTQSPSTLSASVGDRVTITCRASQSISAFLAWYQQKPGKAPNLVIYKClone PA15P1E01 - light chainASSLDSGVPSTFSGSGSGTEYTLTISSLQPDDFATYYCQQYFSSPPTFGQ variable regionGTKVEMK  195 QSISAF Clone PA15P1E01 - CDR-L1  196 KASClone PA15P1E01 - CDR-L2  197 QQYFSSPPT Clone PA15P1E01 - CDR-L3  698EVRLVESGGGLIQPGGSLRLSCAASGFNVSSDYMNWVRQAPGKGLEWVSVClone PA15P1E02 - heavy chainLYSSGFTYYADSVRGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARVAL variable regionFGEPLVDSWGQGTLVTVSS  699 GFNVSSDY Clone PA15P1E02 - CDR-H1  700 LYSSGFTClone PA15P1E02 - CDR-H2  701 ARVALFGEPLVDS Clone PA15P1E02 - CDR-H3 702 EIVMTQSPATLSVSPGERATLSCRASQSVSSNLAWYQQKPGQAPRLLIYGClone PA15P1E02 - light chainAISRATGIPARFSGSGSGTEFTLTISSLQSEDFAIYYCQQYNNWPWTFGQ variable regionGTKVEIK  482 QSVSSN Clone PA15P1E02 - CDR-L1  703 GAIClone PA15P1E02 - CDR-L2  584 QQYNNWPWT Clone PA15P1E02 - CDR-L3  704DPYSPS Binding motif sequence for PA13P1H08  705 DSYGRDPYSPSBinding motif sequence for PA13P1H08  706 YSPSQDPYSPSBinding motif sequence for PA13P1H08  707 PDRRDPYSPSBinding motif sequence for PA13P1H08  708RQQWELQGDRRCQSQLERANLRPCEQHLMQKIQRDEDSYGRDPYSPSAra h 2 isofornn Ara h 2.0201QDPYSPSQDPDRRDPYSPSPYDRRGAGSSQHQERCCNELNEFENNQRCMC(DPYSPS motif underlined)EALQQIMENQSDRLQGRQQEQQFKRELRNLPQQCGLRAPQRCDLEVESGG RDRY  709EVQLVASGGGLIHPGGSLRLSCEASGFSFSRFWMYWVRQSPGEGLVWVARClone 1003320101_D6 heavyLSGDGTVTNYADSMEGRVTISRDNVKNTLFLEMNSLREGDTGIYYCARKDchain variable region sequence CPSLSCQLDYWGQGVQVTVSS  710 GFSFSRFWClone 1003320101_D6 CDR-H1  711 LSGDGTVT Clone 1003320101_D6 CDR-H2  712ARKDCPSLSCQLDY Clone 1003320101_D6 CDR-H3  713QSVLTQPPSVSAAPGQKVTISCSGSTSNIGKNYVSWYQHFPGAAPKLLIFClone 1003320101_D6 light chainDNDKRPSGIPDRFSGSRSGTSATLDITGLQTGDEADYFCATWDSVFGSGTvariable region sequence TVSVL  714 TSNIGKNY Clone 1003320101_D6 CDR-L1 715 DND Clone 1003320101_D6 CDR-L2  716 ATWDSRLSADVClone 1003320101_D6 CDR-L3  717EVQLLESGGGLVQPGGSLRLSCAASGFNFSNFAVSWVRQTPGKGLEWVSAClone 1003320105_D6 heavy chainILGSRSVTYYADSVKGRFTISRDKSKNALYLQMDSLRAEDTAIYYCAKLFvariable region sequence FMPYSHDDSGDYWGQGTLVAVSS  718 GFNFSNFAClone 1003320105_D6 CDR-H1  719 ILGSRSVT Clone 1003320105_D6 CDR-H2  720AKLFFMPYSHDDSGDY Clone 1003320105_D6 CDR-H3  721QLVLTQSPSASASLGASVKLTCTLSSDHRSYAIAWHQQQPGKGPRYLMKVClone 1003320105_D6 light chainNRDGSHIKGDGIPHRFSGSSSVTERYLIISSLQSEDEADYYCQSWDTGIQvariable region sequence VFGGGTRLTVV  722 SDHRSYAClone 1003320105_D6 CDR-L1  723 VNRDGSH Clone 1003320105_D6 CDR-L2  724QSWDTGIQV Clone 1003320105_D6 CDR-L3  725QVQLQESGLGLVKPSGTLSLTCAVSGGPMNSSYWWSWVRQSPGGGLEWIGClone 1003320107_C5 heavy chainQISHYTNTKYNPSFKNRVSISIDKSKNEFSLRLTYVTGADTGVYYCVGERvariable region sequence DWKDPNWFDPWGQGRLVTVSS  726 GGPMNSSYWClone 1003320107_C5 CDR-H1  727 ISHYTNT Clone 1003320107_C5 CDR-H2  728VGERDWKDPNWFDP Clone 1003320107_C5 CDR-H3  729QSVLTQPPSVSGAPGQRVTISCTGSNSNIGAGQDVHWYQHFPGTAPKLVIClone 1003320107_C5 light chainYGNSNRPSGVPDRFSGSKSGTSASLAISGLQADDEADYYCQSYDKSLSSS variable regionLFGGGTKLTVL  730 NSNIGAGQD Clone 1003320107_C5 CDR-L1  731 GNSClone 1003320107_C5 CDR-L2  732 QSYDKSLSSSL Clone 1003320107_C5 CDR-L3 733 QVQLQESGPGLVKPSETLSLSCNVSGGSIRGHYWSWIRQSPGKRLEWLGYClone 1003320107_F3 heavy chainIYQSGYTKYNPSLKSRVSISLDTSKNKFSLNLKSVTTADTAVYYCAGRVAvariable region sequence ERGGDQFDFWGQGTLVTVSS  734 GGSIRGHYClone 1003320107_F3 CDR-H1  735 IYQSGYT Clone 1003320107_F3 CDR-H2  736AGRVAERGGDQFDF Clone 1003320107_F3 CDR-H3  737SYELTQSPSLSVSPGQTASITCSGENLGEKHASWYQQKSGQSPVLVIYQDClone 1003320107_F3 light chainTKRPAGIPERFSGSNSGSTATLTISGTQPMDEADYFCQAWDANTANVIFGvariable region sequence GGTMLTVL  738 NLGEKH Clone 1003320107_F3 CDR-L1 739 QDT Clone 1003320107_F3 CDR-L2  740 QAWDANTANVIClone 1003320107_F3 CDR-L3  741EVQLVESGGGLVQPGGSLKLSCAASGFTFSGSTIHWVRQTSGKGLEWVGRClone 1003320107_F8 heavy chainIGSKATSYATAYAASVKGRFTISRDDSKNTAYLQMNSLKTEDTAVYFCTRvariable region sequence RYYDTTKSVLVVSDSWGQGTLVTVSS  742 GFTFSGSTClone 1003320107_F8 CDR-H1  743 IGSKATSYAT Clone 1003320107_F8 CDR-H2 744 TRRYYDTTKSVLVVSDS Clone 1003320107_F8 CDR-H3  745SYELTQPPSMSVSPGQTARITCSGDVLAKQFAYWYQQKPGQAPVLVIYKDClone 1003320107_F8 light chainSERPSGIPERFSGSSSGTIITLTISGVQAEDEADYYCQSADSSGTSWVFGvariable region sequence GGTKLTVL  746 VLAKQF Clone 1003320107_F8 CDR-L1 747 KDS Clone 1003320107_F8 CDR-L2  748 QSADSSGTSWVClone 1003320107_F8 CDR-L3  749QLLLQESGPGLVKPSETLSLSCTVSAGSITSINYSWGWIRQPPGKGLEWIClone PA01P2C05 heavy chainASVYFSGSIYYNPSLKSRVAISVDTSKNTFSLNLTSVTAADTAVYYCARLvariable region sequence RLDTGRDSSGLSYREHFDYWAQGTLVTVSS  750 AGSITSINYSClone PA01P2C05 CDR-H1  751 VYFSGSI Clone PA01P2C05 CDR-H2  752ARLRLDTGRDSSGLSYREHFDY Clone PA01P2C05 CDR-H3  753DIQMTQSPSTLSASVGDRVTITCRASQSIGMWLAWFQQKPGKAPKLLIYKClone PA01P2C05 light chainASTLESGVPSRFSGSGSGTEFTLTINSLQPDDFATYYCQQYNSYLFTFGPvariable region sequence GTKVDIK  754 QSIGMW Clone PA01P2C05 CDR-L1  196KAS Clone PA01P2C05 CDR-L2  755 QQYNSYLFT Clone PA01P2C05 CDR-L3  756EVQLVQSGAEVKKPGESLKISCKGSGYNFTSSWIGWVRQMPGKGLEWMGIClone PA01P2B03 heavy chainIHPGDSDTRYSPSFQGQVTISADKSLTTAFLQWSSLKTSDTAIYYCARHGvariable region sequence STMLWGDAFDIWGQGTMVTVSS  757 GYNFTSSWClone PA01P2B03 CDR-H1  758 IHPGDSDT Clone PA01P2B03 CDR-H2  759ARHGSTMLWGDAFDI Clone PA01P2B03 CDR-H3  760SYELTQPPSVSLSPGQTARITCSGDALPKHYAYWYQQKPGQAPVLVIYKDClone PA01P2B03 light chainTERPSGIPERFSGSSSGTTVTLTISGVQAEDEADYYCQSSDSTGEVFGGGvariable region sequence TKLTVL  761 ALPKHY Clone PA01P2B03 CDR-L1  762KDT Clone PA01P2B03 CDR-L2  763 QSSDSTGEV Clone PA01P2B03 CDR-L3  764QVQLVQSGAEVKKPGASVMLSCKASGYIFTNSDINWVRQAPGQGPEWMGWClone PA01P2A12 heavy chainMNPKSGNTGYEQKFQGRVTMTTNTSISTAYMELSRLRSEDTAVYYCARSTvariable region sequence GAVAGNFDYWGQGTPVTVSS  765 GYIFTNSDClone PA01P2A12 CDR-H1  766 MNPKSGNT Clone PA01P2A12 CDR-H2  767ARSTGAVAGNFDY Clone PA01P2A12 CDR-H3  768EIVMTQSPATLSVSLGDRATLSCRASQSISRNLAWYQQKPGQAPRLLIYGClone PA01P2A12 light chainASIRITDIPARFSGSGSGTEFTLTISSLQSEDFAIYFCQQYNNWRTFGQGvariable region sequence TRVELK  769 QSISRN Clone PA01P2A12 CDR-L1   30GAS Clone PA01P2A12 CDR-L2  770 QQYNNWRT Clone PA01P2A12 CDR-L3  771HVQLQESGPGLVKSSETLSLTCNVSSDSFSDHYWSWVRQPAGKGLQWLGRClone PA01P2C12 heavy chainIYNTGTTTYNPSLNRRITMSVDTSKNQFSLRLTSVTAADTAVYYCAARHYvariable region sequence HYDKTIWGQGTLVTVSS  772 SDSFSDHYClone PA01P2C12 CDR-H1  773 IYNTGTT Clone PA01P2C12 CDR-H2  774AARHYHYDKTI Clone PA01P2C12 CDR-H3  775NFMLTQPHSVSESPGKTVTISCTRSSGSIASNYVQWYQQRPGSPPTTVIYClone PA01P2C12 light chainEDNQRPSGVPDRFSGSIDSSSNSASLTISGLKPEDEADFYCQSYDSDDREvariable region sequence VFGGGTRLTVL  776 SGSIASNYClone PA01P2C12 CDR-L1  389 EDN Clone PA01P2C12 CDR-L2  777 QSYDSDDREVClone PA01P2C12 CDR-L3  778QVQLVQSGAEVKKPGASVKVSCKASGYTFTTYGISWVRQAPGQGLEWMGWISAYNGNTTYAQNFHARVTMTTDTSTSTAYMELRSLRSDDTAVYFCARTSClone PA01P2E10 heavy chain ARTITIFGVLIPAGLNLDYWGQGTLVTVSSvariable region sequence  779 GYTFTTYG Clone PA01P2E10 CDR-H1  532ISAYNGNT Clone PA01P2E10 CDR-H2  780 ARTSARTITIFGVLIPAGLNLDYClone PA01P2E10 CDR-H3  781QSALTQPASVSGSPGQSITISCTGIGSDVGRYNYVSWYQQHPGKAPKFMIClone PA01P2E10 light chainYDVSNRPSGVSNRFSASKSGNTASLTISGLQAEDEADYYCSSYTSTSTLVvariable region sequence FGGGTKLTVL  782 GSDVGRYNYClone PA01P2E10 CDR-L1  404 DVS Clone PA01P2E10 CDR-L2  783 SSYTSTSTLVClone PA01P2E10 CDR-L3  784QVDLVESGGGYVKSGGSLRLSCAASGFRFSDYYMSWVRQAPGKGLEWLSHISSDSSDTNYADSVKGRFSISRDNAKNSVFLQMNTLRAEDTAVYYCARNAClone PA01P2C09 heavy chain LTNAYDMSGFRNWGQGILVTVSSvariable region sequence  785 GFRFSDYY Clone PA01P2C09 CDR-H1  786ISSDSSDT Clone PA01P2C09 CDR-H2  787 ARNALTNAYDMSGFRNClone PA01P2C09 CDR-H3  788NFMLTQPHSVSESPGKTVILSCTRSSGSIATNYVRWYQQRPGSAPTTVIYClone PA01P2C09 light chainEDSRRPSSVPDRFSGSIDSSSNSASLTISGLRTEDEADYYCQSFDTSSRKvariable region sequence VVFGGGTKLTVL  789 SGSIATNYClone PA01P2C09 CDR-L1  790 EDS Clone PA01P2C09 CDR-L2  791 QSFDTSSRKVVClone PA01P2C09 CDR-L3  792QVTLRESGPALVEVTQTVTLTCNFSGFSLHTRGMYVNWIRQPPGKALEWLAVINWDDDKYYTPSLKNRLTISKDTSKNQVVLTMTNMDPVDTATYYCARTClone PA01P1D06 heavy chain DYGGYGPEGFDYWGQGTLVTVSSvariable region sequence  793 GFSLHTRGMY Clone PA01P1D06 CDR-H1  794INWDDDK Clone PA01P1D06 CDR-H2  795 ARTDYGGYGPEGFDYClone PA01P1D06 CDR-H3  796EIVMTQSPATLSVSPGESATLSCRASQSVRSNLAWYQQKPGQAPRLLIYGClone PA01P1D06 light chainTASTRAPGVPARFGSESGREFTLTISSLQSEDFAVYYCQQYNNWPPYTFGvariable region sequence QGTKLEIK  797 QSVRSN Clone PA01P1D06 CDR-L1  30 GAS Clone PA01P1D06 CDR-L2  798 QQYNNWPPYT Clone PA01P1D06 CDR-L3 799 QVQLVESGGGVVQPGRSLRLSCVASGFDLNAYGMHWVRQAPGKGLDWVAAClone PA01P2E08 heavy chainTSRGGTKKYYADSVKGRFTISRDVSKNTLYLQMGSLRTGDTAIYYCGVGMvariable region sequence EDVFDIWGQGTMVTVSP  800 GFDLNAYGClone PA01P2E08 CDR-H1  801 TSRGGTKK Clone PA01P2E08 CDR-H2  802GVGMEDVFDI Clone PA01P2E08 CDR-H3  803QSVLTQPPSVSAAPGQKVTISCSENNSNIGNRNVSWYQQLPGKAPKLFIYClone PA01P2E08 light chainDNNERPSGIPARFSGSKSGTSATLVITGLQTGDEADYYCGTWDRSLSVWVvariable region sequence FGGGTKLTVL  804 NSNIGNRN Clone PA01P2E08 CDR-L1 110 DNN Clone PA01P2E08 CDR-L2  805 GTWDRSLSVWV Clone PA01P2E08 CDR-L3 806 QVQLQESGPGLVKPSQTLSLTCTVSGGSMRSGDYYWSWIRQPPGKGLEWIClone PA01P2A05 heavy chainGYIYFTGSSYYNPSLKSRATISVDTSKNQFSLKLNSVTAADTAVYFCARGvariable region sequence VDVDLTFFDCWGHGTLVTVSS  807 GGSMRSGDYYClone PA01P2A05 CDR-H1  808 IYFTGSS Clone PA01P2A05 CDR-H2  809ARGVDVDLTFFDC Clone PA01P2A05 CDR-H3  810SYVLTQPPSVSLAPGKTARITCGGNNIGNKSVHWYQQKPGQAPVLVIYYDClone PA01P2A05 light chainSDRPSGIPERFSGSNSGNTATLTINRVEAGDEADYHCQVWDSSTDHRVFGvariable region sequence EGTKLTVL  811 NIGNKS Clone PA01P2A05 CDR-L1 509 YDS Clone PA01P2A05 CDR-L2  812 QVWDSSTDHRV Clone PA01P2A05 CDR-L3 813 QVQLQQWGAGLLKPSETLSLTCTVIGTSFSNYYWSWIRQPPGKGLQWIGEClone PA01P2B04 heavy chainITHSDSANYNPSLKSRVIISIDSSKNQLSLNLSSVTAADTAVYYCARGSKvariable region sequence DYYDRSTFSWFDPWGQGTLVTVSS  814 GTSFSNYYClone PA01P2B04 CDR-H1  815 ITHSDSA Clone PA01P2B04 CDR-H2  816ARGSKDYYDRSTFSWFDP Clone PA01P2B04 CDR-H3  817EIVMTQSPATLSVSPGERATLSCRASQNISNKLAWYQQKPGQAPRLLIYDClone PA01P2B04 light chainASTRATGVPARFSCSVSGTAFTLTINRLQSEDFAVYYCQQYYYWPPPYTFvariable region sequence GHGTKLEIK  818 QNISNK Clone PA01P2B04 CDR-L1  94 DAS Clone PA01P2B04 CDR-L2  819 QQYYYWPPPYT Clone PA01P2B04 CDR-L3 820 QVQLVESGGGFVKPGGSLRLSCAVSGFTFSDYYMSWVRQAPGKGLEWLSHClone PA01P2E05 heavy chainISSDGSDTNYADSVKGRFSISRDNAKNSVFLQMNTLRVEDTAVYYCARNAvariable region sequence LTNAYDMSGFRNWGQGTLVTVSS  821 GFTFSDYYClone PA01P2E05 CDR-H1  822 ISSDGSDT Clone PA01P2E05 CDR-H2  787ARNALTNAYDMSGFRN Clone PA01P2E05 CDR-H3  823NFMLTQPHSVSESPGKTVILSCTRSSGSIASNYVRWYQQRPGSAPTTVIYClone PA01P2E05 light chainEDSRRPSSVPDRFSGSIDSSSNSASLTISGLKTEDEADYYCQSFDSSSRKvariable region sequence VVFGGGTKLTVL  776 SGSIASNYClone PA01P2E05 CDR-L1  790 EDS Clone PA01P2E05 CDR-L2  824 QSFDSSSRKVVClone PA01P2E05 CDR-L3  825QVQLLQSGPEVKQPGASVQVSCQTSGYTFTGYYIHWVRQAPGQGLEWVGWClone PA01P2D04 heavy chainINPNRGHTNYGPAFQGRLTLTADTSSSTAYLELTRLRSDDTAVYYCARDRvariable region sequence LTGGRDAFEIWGQGTMLIVSS  121 GYTFTGYYClone PA01P2D04 CDR-H1  826 INPNRGHT Clone PA01P2D04 CDR-H2  827ARDRLTGGRDAFEI Clone PA01P2D04 CDR-H3  828DIQLTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLISVClone PA01P2D04 light chainASSLQDGVPPRFSGSRSGTEFTLTISSLQPEDFAIYYCQQSYSLSWTFGQvariable region sequence GTKVEIK  829 QSISSY Clone PA01P2D04 CDR-L1  830VAS Clone PA01P2D04 CDR-L2  831 QQSYSLSWT Clone PA01P2D04 CDR-L3  832QVQLVQSGAEVKQPGASVQVACQTSGYIFTAYYIHWLRQAPGQGLEWVGWClone PA01P2B12 heavy chainINPNRGHTNYAPGFQGRLTLTADTSSSTAYLALTRLASDDTAVYYCARDRvariable region sequence LTGGRDAFEIWGQGTMLIVSS  833 GYIFTAYYClone PA01P2B12 CDR-H1  826 INPNRGHT Clone PA01P2B12 CDR-H2  827ARDRLTGGRDAFEI Clone PA01P2B12 CDR-H3  834DIQLTQSPSSLSASVGDRVTITCRASQSVSSYLNWYQQKPGEAPKLLISAClone PA01P2B12 light chainASSLQDGVPPRFSGSRSGTEFTLTISSLQPEDFAIYYCQQSYSLWWTFGQvariable region sequence GTKVEIK  835 QSVSSY Clone PA01P2B12 CDR-L1  149AAS Clone PA01P2B12 CDR-L2  836 QQSYSLWWT Clone PA01P2B12 CDR-L3  837QLQLQESGSGLVKPSQTLSLTCDVSGDSMNDDVYTWSWIRQPPGRGLEWIClone PA01P2D11 heavy chainGYISHTGNTFYNSSLQSRVTMSVDTSKNQFSLKLSSVTIADTAVYYCARLvariable region sequence TFLFSAPFSSFNWFDPWGQGILVTVSS  838 GDSMNDDVYTClone PA01P2D11 CDR-H1  839 ISHTGNT Clone PA01P2D11 CDR-H2  840ARLTFLFSAPFSSFNWFDP Clone PA01P2D11 CDR-H3  841QSVLTQPPSVSGAPGQTITISCTGTPSNFGADYDVHWYQQRPGTAPKLLIClone PA01P2D11 light chainFADKHRPSGVPDRFSGSRSGTSASLAISGLQAEDEADYYCQSYDSGVVGLvariable region sequence WVFGGGTKVTVL  842 PSNFGADYDClone PA01P2D11 CDR-L1  843 ADK Clone PA01P2D11 CDR-L2  844 QSYDSGVVGLWVClone PA01P2D11 CDR-L3  845QVQLQQWGAGLLKPSETLSLTCGVHGGSLNNYYWSWIRQPPGKGLEWIGEClone PA01P2B10 heavy chainVYHSGSINYNPSLKSRVTMSVDTSKNQFSFNLSSVTAADTAVYYCARGAYvariable region sequence DSRGFWILDAFNTWGQGTMVIVSS  846 GGSLNNYYClone PA01P2B10 CDR-H1  847 VYHSGSI Clone PA01P2B10 CDR-H2  848ARGAYDSRGFWTLDAFNT Clone PA01P2B10 CDR-H3  849DIQMTQSPSALSASLGDRVTITCRASESINSWLAWYQQKPGKAPKLLIYKClone PA01P2B10 light chainASTLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCHQYNRYSYTFGQvariable region sequence GTKLDIK  850 ESINSW Clone PA01P2B10 CDR-L1  196KAS Clone PA01P2B10 CDR-L2  851 HQYNRYSYT Clone PA01P2B10 CDR-L3  852EVLLLESGGGLVHPGGTLRLSCAASGFTFRNSAMTWVRQAPGKGLEWVSSClone PA01P2D10 heavy chainIGGSGAKSYYADSVKGRFTISRDNSKNTLYLEMNTLRVDDTAIYYCAKDQvariable region sequence LNCYDLWSGDYCWFDTWGQGTLVTVSS  853 GFTFRNSAClone PA01P2D10 CDR-H1  854 IGGSGAKS Clone PA01P2D10 CDR-H2  855AKDQLNCYDLWSGDYCWFDT Clone PA01P2D10 CDR-H3  856QSVLIQPPSASGTPGQRVTISCSGSNSNIGSNYVCWYQHLPGGAPKLLIYClone PA01P2D10 light chainRNNQRPSGVPDRFSGSKSGTSASLAISGLRSEDEADYYCAAWDDSLSGSWvariable region sequence VFGGGTKLTVL  857 NSNIGSNYClone PA01P2D10 CDR-L1  662 RNN Clone PA01P2D10 CDR-L2  858 AAWDDSLSGSWVClone PA01P2D10 CDR-L3  859EVQLLESGGGLVQPGGSLRLSCAVSGLKFSSYAMSWVRQAPGKGLEWVSVClone PA01P2D09 heavy chainVSGSSGSTFYAVSVEGRFSISRDNSNNMLYMDMHSLRVEDTAKYYCAKVVvariable region sequence GWYYDRNGNRRPKGFRAFDVWGQGTMVIVSS  860 GLKFSSYAClone PA01P2D09 CDR-H1  861 VSGSSGST Clone PA01P2D09 CDR-H2  862AKVVGWYYDRNGNRRPKGFRAFDV Clone PA01P2D09 CDR-H3  863QSVLTQPPSASGTPGQRITIACSGTTSNIGGNSVNWYQQFPGAAPRLLIFClone PA01P2D09 light chainDYDQRPSGVPARFSGSSSGSSGYLAISGLQSEDEADYYCSSWDDNLNGWVvariable region sequence FGGGTKLTVL  864 TSNIGGNS Clone PA01P2D09 CDR-L1 865 DYD Clone PA01P2D09 CDR-L2  866 SSWDDNLNGWV Clone PA01P2D09 CDR-L3 867 EVQLLESGGGLVQPGGSLRLSCVASGFTFSGHAMSWVRQAPGKGLEWVSGClone PA01P2B05 heavy chainISGSGGSTYYADSVKGRFTISRDNSKKTVDLQMNNLRAEDTAIYYCAKDLvariable region sequence HFDTSGYYYSMIFDYWGQGTLVPVSS  868 GFTFSGHAClone PA01P2B05 CDR-H1  378 ISGSGGST Clone PA01P2B05 CDR-H2  869AKDLHFDTSGYYYSMIFDY Clone PA01P2B05 CDR-H3  870QSALAQPASVSGSPGQSITISCTGTSSDISDYNYVSWYQQHPGKAPKLILClone PA01P2B05 light chainYDVNNRPSGVSSRFSGSKSGDTASLTISGLQPEDEADYYCSSYTSTKIFGvariable region sequence GGTKVTVL  871 SSDISDYNY Clone PA01P2B05 CDR-L1 682 DVN Clone PA01P2B05 CDR-L2  872 SSYTSTKI Clone PA01P2B05 CDR-L3 873 EVQLLESGGGLLQLGGSLRLSCAASGFTFSSYVMSWVRQAPGKGLEWVSLClone PA01P4C11 heavy chainITGSGGNTYYADSVKGRFTISRDNSKNTLFLQMNSLRVEDTAIYYCVKTDvariable region sequence FYDSSGYYFHDAFHIWGQGTMVTVSS  874 GFTFSSYVClone PA01P4C11 CDR-H1  875 ITGSGGNT Clone PA01P4C11 CDR-H2  876VKTDFYDSSGYYFHDAFHI Clone PA01P4C11 CDR-H3  877QTVVTQEPSLTVSPGGTVTLTCASSTGSVTSGYYPNWFQQKPGQAPRTLIClone PA01P4C11 light chainYGTSNKHSWTPARFSGSLLGGKAALTLSDVQPEDEAEYYCLLYYGGAYVFvariable region sequence GTGTKVTVL  878 TGSVTSGYY Clone PA01P4C11 CDR-L1  22 GTS Clone PA01P4C11 CDR-L2  879 LLYYGGAYV Clone PA01P4C11 CDR-L3 880 QVQLVQSGAEVKKPGASVKVSCKASGYTFIRYDIHWVRQATGQGLEWMGWClone PA01P3E08 heavy chainMNPNNGKSGFAQKFEGRVTLTRNTSVTSTYMQLSSLGLEDTAVYYCVRAGvariable region sequence YSYGWGFDYWGQGSLVTVSS  881 GYTFIRYDClone PA01P3E08 CDR-H1  882 MNPNNGKS Clone PA01P3E08 CDR-H2  883VRAGYSYGWGFDY Clone PA01P3E08 CDR-H3  884NFTLTQPHSVSGSPGKTVTISCTRSSGGIASSHVQWYQQRPASAPTTLIFClone PA01P3E08 light chainEDDQRSSGVPDRFSGSIDTSSNSAYLTISGLEAEDEADYYCQSYDNSMWVvariable region sequence FGGGSKVTVL  885 SGGIASSH Clone PA01P3E08 CDR-L1 886 EDD Clone PA01P3E08 CDR-L2  887 QSYDNSMWV Clone PA01P3E08 CDR-L3 888 HVQLVQSGADVKKPGSSVKVSCKFSGGTFNNDSINWVRQAPGQGLEWMGVClone PA01P2E06 heavy chainIMPFFGATRFAPKFQGRVTLTADKFTSTGYMELGSLKSDDTAVYYCARDKvariable region sequence PPDDKWADYGMDVWGQGTTVTVSS  889 GGTFNNDSClone PA01P2E06 CDR-H1  890 IMPFFGAT Clone PA01P2E06 CDR-H2  891ARDKPPDDKWADYGMDV Clone PA01P2E06 CDR-H3  892SYELTQPPSVSVSPGQTARITCSGDALPKQYVYWYQQKTGQAPVLVIYKDClone PA01P2E06 light chainTERPSGIPERFSGSTSGTTVTLTISGVQADDEADYFCQSADRNANYRVFGvariable region sequence GGTKLTVL  523 ALPKQY Clone PA01P2E06 CDR-L1 762 KDT Clone PA01P2E06 CDR-L2  893 QSADRNANYRV Clone PA01P2E06 CDR-L3 894 QLQLQESGSGLVKPSQTLSLTCAVSGGSITSGTYSWTWIRQSPEKGLEWIClone PA01P2E07 heavy chainGYIYYTGSTYYNPSLGRRVTISGDTSNNEFSLNLKSVTAADTAVYYCARGvariable region sequence IHRGGVLDFWGQGILVTVSS  895 GGSITSGTYSClone PA01P2E07 CDR-H1  896 IYYTGST Clone PA01P2E07 CDR-H2  897ARGIHRGGVLDF Clone PA01P2E07 CDR-H3  898EIVLTQSPATLPLSPGERATLSCRASQSLDKYLAWYQQKPGQAPRLLIYDClone PA01P2E07 light chainTSKRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYFCQQRNNWPPYTFGvariable region sequence QGTKVEMK  899 QSLDKY Clone PA01P2E07 CDR-L1 900 DTS Clone PA01P2E07 CDR-L2  901 QQRNNWPPYT Clone PA01P2E07 CDR-L3 902 QVLLVQSGSEVKNPGASIRVSCKTSGYMFTNNGIAWVREVPTQGLEWMGWClone PA01P2G07 heavy chainISTYSGATHYAPNLHGRITMTADTSASTAYMELRSLQSGDTGVYYCARLWvariable region sequence FGKLGLDFWGQGTQVTVSS  903 GYMFTNNGClone PA01P2G07 CDR-H1  904 ISTYSGAT Clone PA01P2G07 CDR-H2  905ARLWFGKLGLDF Clone PA01P2G07 CDR-H3  906QSVLTQPPSASGTPGQRVIISCSGSTSNIGTKTVNWYQHLPGTAPKLLIYClone PA01P2G07 light chainNNNQRPSGVPDRFSGSKSGTSASLTISGLQSEDEADYYCAAWDDSLNGRGvariable region sequence LFGPGTKVTVL  907 TSNIGTKTClone PA01P2G07 CDR-L1  908 NNN Clone PA01P2G07 CDR-L2  909 AAWDDSLNGRGLClone PA01P2G07 CDR-L3  910QVEVVESGGGVVQPGKSLRLSCAASGFKFNVYGIHWVRQAPGKGLEWVAVClone PA01P2B09 heavy chainVWYDGSNKYYADSVKGRFTISRDNSKNTTYLQMDSLRVDDTAVYYCARELvariable region sequence QYSNYDYFYAMDVWGQGTTVTVSS  911 GFKFNVYGClone PA01P2B09 CDR-H1  912 VWYDGSNK Clone PA01P2B09 CDR-H2  913ARELQYSNYDYFYAMDV Clone PA01P2B09 CDR-H3  914DIQMTQSPPSLSASVGDRVTITCRASQDIDNYLVWFQQKPGRAPKSLIYAClone PA01P2B09 light chainASSLQSGVPSKFSGSGSGTEFTLTISSLQPEDFATYYCQQYNSFPYTFGQvariable region sequence GTKLEIK  915 QDIDNY Clone PA01P2B09 CDR-L1  149AAS Clone PA01P2B09 CDR-L2  916 QQYNSFPYT Clone PA01P2B09 CDR-L3  917QVQLQESGPGLVKPSETLSLTCSVSGGSISSHYWSWIRQPPGRGLEWIAYClone PA01P2C04 heavy chainISYSGRTKYNPSLKSRVTISEDTSKNQFSLKLSSVTPADTAVYYCARIYGvariable region sequence DYGPFIDYWGQGTLVTVSS  918 GGSISSHYClone PA01P2C04 CDR-H1  919 ISYSGRT Clone PA01P2C04 CDR-H2  920ARIYGDYGPFIDY Clone PA01P2C04 CDR-H3  921DIQMTQSPSSLSASVGDRVTITCRASQTISTYLNWYQQKPGTAPMLLIYGClone PA01P2C04 light chainAYSLHSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSSSLPLTFGGvariable region sequence GTKVEIK  922 QTISTY Clone PA01P2C04 CDR-L1  923GAY Clone PA01P2C04 CDR-L2  924 QQSSSLPLT Clone PA01P2C04 CDR-L3  925QEQLQESGPGLVKPSQTLSLTCTVSGGSISSGDHYWSWLRQTPGKGLEWIClone PA01P2H08 heavy chainGYIYYRGNTNYNPSLESRITMSVDTSKNQFSLKLSSVTAADTGVYYCARDvariable region sequence RRLLFWFGQGPETFDIWGPGTMVTVSS  926 GGSISSGDHYClone PA01P2H08 CDR-H1  927 IYYRGNT Clone PA01P2H08 CDR-H2  928ARDRRLLFWFGQGPETFDI Clone PA01P2H08 CDR-H3  929DIQMTQSPSILSASVGDRVTITCRASQNINHWLAWYQQKPGKAPKLLIYMClone PA01P2H08 light chainASSLENGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQYNSYSGTFGHvariable region sequence GTKVDIK  930 QNINHW Clone PA01P2H08 CDR-L1  931MAS Clone PA01P2H08 CDR-L2  932 QQYNSYSGT Clone PA01P2H08 CDR-L3  933TACTGTGCGAAAGTTCTTGACTACAGTGAATTTCATTACTATTACGGTTT FIG. 13A PA12P3F10GGACGTCTGGGGCCAAGGGACCGCGGTCGCCGTCTCCTCAG  934TACTGTGCGAAAGTTCTTGACTACAATGAGTACTCTCTCTACTTCGGTAT FIG. 13A PA12P3D08GGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCAG  935TACTGTGCGAAAGTTCTTGACTACAGTGAATACTCTCTCTACTTCGGTAT FIG. 13A PA12P1C07GGACGTCTGGGGCCAAGGGACCACGGTCCTTGTCTCCTCAG  936TACTGTGCGAAGGTCCTTGACTACAGTAGGTACTCCTATTATTACGGGAT FIG. 13A PA13P1H08GGACGTCTGGGGCCAGGGGACCACGGTCATCGTCTCCTCAG  937TACTGTGCTAAGGTCCTTGACTACAGTGCATTCTCCTATTATTATGGGAT FIG. 13A PA13P1E10GGACGTCTGGGGCCAGGGGACCACGGTCATCGTCTCCTCAG  938TATTGTGCGAAAGTCCTTGACTACAGTATTTTCTATTACTATTTCGGCCT FIG. 13A PA13P3G09GGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCAG  939 TACTGTGCGAAAGAFIG. 13A IGHV30-30*18  940 TYCT FIG. 13A Nontennplated  (inferred)  941TGACTACAGTAACTAC FIG. 13A IGHD4-11*01  942ATTACTACTACTACTACGGTATGGACGTCTGGGGCCAAGGGACCACGGTC FIG. 13A IGHJ6*02ACCGTCTCCTCAG  943 YCAK FIG. 13A Translated: V  944 VLDYSNYFIG. 13A Translated:  Nontennplated + D  945 NYYYYYGMDVWGQGTIVIVSSFIG. 13A Translated: J  946 YCAKVLDYSNYYYYYGMDVWGQGTTVTVSSFIG. 13A Inferred naive rearrangement  947YCAKVLDYSEFHYYYGLDVWGQGTAVAVSS FIG. 13A Translated: PA12P3F10  948YCAKVLDYNEYSLYFGMDVWGQGTTVTVSS FIG. 13A Translated: PA12P3D08  949YCAKVLDYSEYSLYFGMDVWGQGTTVLVSS FIG. 13A Translated: PA12P1C07  950YCAKVLDYSRYSYYYGMDVWGQGTTVIVSS FIG. 13A Translated: PA13P1H08  951YCAKVLDYSAFSYYYGMDVWGQGTIVIVSS FIG. 13A Translated: PA13P1E10  952YCAKVLDYSIFYYYFGLDVWGQGTTVTVSS FIG. 13A Translated: PA13P3G09  953GCAGTGTATTACTGTCAGCATTACAGTAATTCACCCCCGTACACTTTTGG FIG. 13C PA12P3F10CCCGGGGACCAAGTTGGAGATCAAAC  954GCAGTGTATTTCTGTCAGTACTATAGTGACTCACCTCCGTACACTTTTGG FIG. 13C PA12P3D08CCCGGGGACCAAGCTGGAGATCAAAC  955GCAGTGTATTCCTGTCAACACTATAGTGACTCACCTCCTTACACTTTTGG FIG. 13C PA12P1C07CCAGGGGACCAAACTGGAGATCAAAC  956GCAGTTTATTACTGTCAGCACTATGGTAGGTCACCTCCGTACACTTTTGG FIG. 13C PA13P1H08CCCGGGGACCAAGCTGGACATCAAAC  957GCAGTATATTACTGTCAACACTATGGTAGGTCACCTCCATACACTTTTGG FIG. 13C PA13P1E10CCAGGGGACCAAAGTGGAGATCAAAC  958GCAGTGTACTACTGTCAGCACTATGGAGACTCACCTCCGTACACCTTTGG FIG. 13C PA13P3G09CCAGGGGACGAAAGTGGAGATGAAAC  959 GCAGTGTATTACTGTCAGCAGTATGGTAGCTCACCTCCFIG. 13C IGKV3-20*01  960 TGTACACTTTTGGCCAGGGGACCAAGCTGGAGATCAAACFIG. 13C IGKJ2*01  961 AVYYCQQYGSSP FIG. 13C Translated: V  962PYTFGQGTKLEIK FIG. 13C Translated: J  963 AVYYCQQYGSSPPYTFGQGTKLEIKFIG. 13C Inferred naive rearrangement  964 AVYYCQHYSNSPPYTFGPGTKLEIKFIG. 13C Translated: PA12P3F10  965 AVYFCQYYSDSPPYTFGPGTKLEIKFIG. 13C Translated: PA12P3D08  966 AVYSCQHYSDSPPYTFGQGTKLEIKFIG. 13C Translated: PA12P1C07  967 AVYYCQHYGRSPPYTFGPGTKLDIKFIG. 13C Translated: PA13P1H08  968 AVYYCQHYGRSPPYTFGQGTKVEIKFIG. 13C Translated: PA13P1E10  969 AVYYCQHYGDSPPYTFGQGTKVEMKFIG. 13C Translated: PA13P3G09  970EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYD FIG. 13E swapASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPGFTF GPGTKVDIK  971AKVMDYDIFKNYFGLDV FIG. 16  972 AKVMDYDVFKNYYGLDV FIG. 16  973AKTLDYSQYMYYYGLDV FIG. 16  974 QHYGRSPPYT FIG. 12G  975 QHYGSSPPFTFIG. 12G  976 QHYGSLPPFT FIG. 12G  977 AAWDDTLVGV FIG. 12G  978AAWDDLVVGV FIG. 12G  979 QSTDSSGDYVV FIG. 12G  980 QSTDSSLRDVV FIG. 12G 981 TSYAGRNIQV FIG. 12G  982 SSYAGSNIAV FIG. 12G  983 QSYDGSSPVIFIG. 12G  984 QSYDTNIVV FIG. 12G  985 QSYDSANVV FIG. 12G  986 QSYDADNAVFIG. 12G  987 SSYTRETALGGV FIG. 12G  988 QQYYTTPRT FIG. 12G  989QQYYTTPYT FIG. 12G  990 QQYLTTPYT FIG. 12G  991 QQYDEWPPFT FIG. 12G  992QQYNHWPPYT FIG. 12G  993 GSYKSGSTWV FIG. 12G  994 SSYRSGSTWV FIG. 12G 995 SSYTSGRTWV FIG. 12G  996 SSYTTGRTWV FIG. 12G  997 ASRYCTDSGCYLGSFDYFIG. 12G  998 ASRYCTDDGCYLGSFDF FIG. 12G  999 TRDHGYY FIG. 12G 1000ARDHGYY FIG. 12G 1001 ARDPAAGTWWFDP FIG. 12G 1002 ARPSAHYYDRGGYNDAFDMFIG. 12G 1003 TTGYRTTTTYHGDDAFDI FIG. 12G 1004 TTGYRTSTSYHGDDAFDIFIG. 12G 1005 ARGPPAVQGYFYYMYV FIG. 12G 1006 ARGPPGVHGYFYYTDV FIG. 12G1007 ARDVVRPGSGPRLGFDP FIG. 12G 1008 ARDVVRPGRGPRLGFDP FIG. 12G 1009AKEGGSSTSWYSLYHEYEMDV FIG. 12G 1010 AHKAAEPGSRDRWFDS FIG. 12G 1011AGGYNNSSFYFDS FIG. 12G 1012 AVGYNNSWFYFDY FIG. 12G 1013 ARLGHLRGWFDSFIG. 12G 1014 VLSQYEFGSSWFYYYRMDV FIG. 12G 1015 VLSKYEFGSSWFYYYRMDVFIG. 12G 1016 VLSKYEFHSSWFYYYRMDV FIG. 12G

Although the foregoing invention has been described in some detail byway of illustration and example for purposes of clarity ofunderstanding, one of skill in the art will appreciate that certainchanges and modifications may be practiced within the scope of theappended claims. In addition, each reference provided herein isincorporated by reference in its entirety to the same extent as if eachreference was individually incorporated by reference.

What is claimed is:
 1. A method of identifying human monoclonalantibodies that specifically bind to an allergen, the method comprising:(a) isolating single B cells from a tissue or body fluid sample; (b)generating cDNA from the single B cells; (c) sequencing the cDNA toidentify (i) a heavy chain variable region sequence of an immunoglobulinheavy chain that comprises an IgE or IgG4 constant region, and (ii) alight chain variable region sequence in an immunoglobulin light chainthat is co-expressed with the immunoglobulin heavy chain in the samesingle B cell; (d) expressing antibodies comprising the heavy and lightchain variable region sequences of step (c); and (e) determining whethersaid antibodies bind said allergen.
 2. The method of claim 1, whereinsaid isolating step comprises sorting cells by fluorescent activatedcell sorting (FACS).
 3. The method of claim 1, wherein said isolatingstep comprises selecting B cells that express cell surface IgE or IgG44. The method of claim 1, wherein said isolating step comprisescontacting cells from the sample with an anti-human CD19 antibody and ananti-human IgE antibody and selecting for CD19+IgE-expressing B cells.5. The method of claim 1, wherein said isolating step comprisescontacting cells from the sample with an anti-human CD19 antibody, ananti-human IgE antibody, an anti-human IgM antibody, and an anti-humanIgG antibody and selecting for CD19+IgM−IgG−IgE-expressing B cells. 6.The method of claim 1, wherein said isolated B cells express a B cellmarker and are negative for non-IgE isotypes.
 7. The method of claim 1,wherein said isolating step comprises contacting cells from the samplewith an anti-human CD19 antibody, an anti-human IgM antibody, ananti-human IgG antibody, an anti-human IgA antibody, and an anti-humanIgD antibody and selecting for CD19+IgM−IgG−IgA−IgD− B cells.
 8. Themethod of claim 1, wherein the isolated B cells express a B cell markerand are negative for non-IgG4 isotypes.
 9. The method of claim 8,wherein said isolating step comprises contacting cells from the samplewith an anti-human CD19 antibody, an anti-human IgM antibody, ananti-human IgE antibody, an anti-human IgA antibody, an anti-human IgDantibody, an anti-human IgG1 antibody, an anti-human IgG2 antibody, andan anti-human IgG3 antibody and selecting forCD19+IgM−IgE−IgA−IgD−IgG1−IgG2−IgG3− B cells.
 10. The method of claim 1,wherein the method further comprises substituting the constant region ofan antibody identified in said sequencing step with a wild-type IgG4constant region or a modified IgG4 constant region sequence.
 11. Themethod of claim 1, wherein the sample is selected from the groupconsisting of blood, sputum, saliva, and tissue biopsy.
 12. The methodof claim 1, wherein the allergen is a food allergen, a plant allergen, afungal allergen, an animal allergen, a drug allergen, a cosmeticallergen, or a latex allergen.
 13. The method of claim 12, wherein theallergen is a food allergen selected from the group consisting of a milkallergen, an egg allergen, a nut allergen, a fish allergen, a shellfishallergen, a soy allergen, a legume allergen, a seed allergen, and awheat allergen.